Isolated binding proteins, e.g., antibodies or antigen binding portions thereof, which bind to tumor necrosis factor-alpha (TNF-α), e.g., human TNF-α, and related antibody-based compositions and molecules are disclosed. Also disclosed are pharmaceutical compositions comprising the antibodies, as well as therapeutic and diagnostic methods for using the antibodies.
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1. A binding protein that binds human TNF-α, the binding protein comprising at least one heavy chain variable region (VH region) and at least one light chain variable region (VL region), wherein
the VH region comprises the amino acid sequence of SEQ ID NO: 74 and
the VL region comprises the amino acid sequence of SEQ ID NO: 84.
2. The binding protein of
3. The binding protein of
(a) a heavy chain constant region comprising an amino acid sequence of SEQ ID NO:2 or SEQ ID NO: 3; and
(b) a light chain constant region comprising an amino acid sequence of SEQ ID NO:4 or SEQ ID NO: 5.
5. The binding protein of
6. The binding protein of
7. A pharmaceutical composition comprising the binding protein of
8. A pharmaceutical composition comprising the binding protein of
9. A pharmaceutical composition comprising the binding protein of
11. A pharmaceutical composition comprising the binding protein of
12. The binding protein of
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This application claims priority to U.S. Provisional Application Ser. No. 61/550,587, filed Oct. 24, 2011, which is incorporated herein by reference in its entirety.
The instant application contains a Sequence Listing which has been submitted electronically in ASCII format and is hereby incorporated by reference in its entirety. Said ASCII copy, created on Jul. 3, 2014, is named 532365_BBI-387_Sequence_Listing.txt and is approximately 1,223,428 bytes in size. This sequence listing replaces a previous sequence listing in ASCII format that was submitted Feb. 27, 2013, and which was also incorporated by reference in its entirety.
Field of the Invention
TNF-α binding proteins and their uses in the prevention and/or treatment of acute and chronic immunological diseases are provided.
Background of the Invention
There is a need in the art for improved binding proteins capable of binding TNF-α (also referred to as tumor necrosis factor, tumor necrosis factor-alpha, tumor necrosis factor-α, TNF, and cachectin). Provided are a novel family of binding proteins, CDR grafted binding proteins, humanized binding proteins, and fragments thereof, capable of binding TNF-α with high affinity and neutralizing TNF-α.
TNF-α binding proteins, or antigen-binding portions thereof, that bind TNF-α are provided. In an embodiment, the antigen binding domain comprises the VH region chosen from any one of SEQ ID NOs: 22, 24, 26, 28, 30, 32, 34-58, 74-83, 94-266, 478-486, 496-675, 738-762, 778-956, 1053-1062, 1073, 1075, and 1077, or one, two, or three CDRs therefrom. In another embodiment, the antigen binding domain comprises the VL region chosen from any one of SEQ ID NOs: 23, 25, 27, 29, 31, 33, 59-73, 84-93, 267-477, 487-495, 676-737, 763-777, 957-1052, 1063-1072, 1074, 1076, and 1078, or one, two, or three CDRs therefrom. In a particular embodiment, the antigen binding domain comprises a VH region and a VL region, for example, wherein the VH region comprises SEQ ID NOs: 22, 24, 26, 28, 30, 32, 34-58, 74-83, 94-266, 478-486, 496-675, 738-762, 778-956, 1053-1062, 1073, 1075, and 1077, or one, two, or three CDRs therefrom, and the VL region comprises SEQ ID NOs: 23, 25, 27, 29, 31, 33, 59-73, 84-93, 267-477, 487-495, 676-737, 763-777, 957-1052, 1063-1072, 1074, 1076, and 1078, or one, two, or three CDRs therefrom.
In an embodiment, the binding protein binds TNF-α. In another embodiment, the binding protein modulates a biological function of TNF-α. In another embodiment, the binding protein neutralizes TNF-α. In yet another embodiment, the binding protein diminishes the ability of TNF-α to bind to its receptor, for example, the binding protein diminishes the ability of pro-human TNF-α, mature-human TNF-α, or truncated-human TNF-α to bind to its receptor. In yet another embodiment, the binding protein reduces one or more TNF-α biological activities selected from: TNF-dependent cytokine production; TNF-dependent cell killing; TNF-dependent inflammation; TNF-dependent bone erosion; and TNF-dependent cartilage damage.
In an embodiment, the binding protein has an on rate constant (Kon) selected from: at least about 102 M−1s−1; at least about 103 M−1s−1; at least about 104 M−1s−1; at least about 105 M−1s−1; and at least about 106 M−1s−1; as measured by surface plasmon resonance. In another embodiment, the binding protein has an off rate constant (Koff) selected from: at most about 10−3 s−1; at most about 10−4 s−1; at most about 10−5 s−1; and at most about 10−6 s−1, as measured by surface plasmon resonance. In yet another embodiment, the binding protein has a dissociation constant (KD) selected from: at most about 10−7 M; at most about 10−8 M; at most about 10−9 M; at most about 10−10 M; at most about 10−11 M; at most about 10−12 M; and at most 10−13 M.
In another aspect, a method for treating a mammal is provided comprising administering to the mammal an effective amount of the pharmaceutical composition disclosed herein. In another embodiment, a method for reducing human TNF-α activity is provided, the method comprising: contacting human TNF-α with the binding protein disclosed herein such that human TNF-α activity is reduced. In another embodiment, provided is a method for reducing human TNF-α activity in a human subject suffering from a disorder in which TNF-α activity is detrimental, the method comprising administering to the human subject the binding protein disclosed herein such that human TNF-α activity in the human subject is reduced. In another embodiment, provided is a method for treating a subject for a disease or a disorder in which TNF-α activity is detrimental, the method comprising administering to the subject the binding protein disclosed herein such that treatment is achieved.
In one embodiment, the method treats diseases involving immune and inflammatory elements, such as autoimmune diseases, particularly those associated with inflammation, including Crohn's disease, psoriasis (including plaque psoriasis), arthritis (including rheumatoid arthritis, psoratic arthritis, osteoarthritis, or juvenile idiopathic arthritis), multiple sclerosis, and ankylosing spondylitis. Therefore, the binding proteins herein may be used to treat these disorders.
Provided are TNF-α binding proteins, or antigen-binding portions thereof, that bind TNF-α, pharmaceutical compositions thereof, as well as nucleic acids, recombinant expression vectors and host cells for making such binding proteins and fragments. Also provided are methods of using the binding proteins disclosed herein to detect human TNF-α, to inhibit human TNF-α either in vitro or in vivo, and to regulate gene expression or TNF-α related functions.
Unless otherwise defined herein, scientific and technical terms used in connection with the present disclosure shall have the meanings that are commonly understood by those of ordinary skill in the art. The meaning and scope of the terms should be clear, however, in the event of any latent ambiguity, definitions provided herein take precedent over any dictionary or extrinsic definition. Further, unless otherwise required by context, singular terms shall include pluralities and plural terms shall include the singular. In this application, the use of “or” means “and/or”, unless stated otherwise. Furthermore, the use of the term “including”, as well as other forms of the term, such as “includes” and “included”, is not limiting. Also, terms such as “element” or “component” encompass both elements and components comprising one unit and elements and components that comprise more than one subunit unless specifically stated otherwise.
Generally, nomenclatures used in connection with, and techniques of, cell and tissue culture, pathology, oncology, molecular biology, immunology, microbiology, genetics and protein and nucleic acid chemistry and hybridization described herein are those well known and commonly used in the art. The methods and techniques of the present disclosure are generally performed according to conventional methods well known in the art and as described in various general and more specific references that are cited and discussed throughout the present specification unless otherwise indicated. See, e.g., Sambrook et al. Molecular Cloning: A Laboratory Manual (2d ed., Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y. (1989)). Enzymatic reactions and purification techniques are performed according to manufacturer's specifications, as commonly accomplished in the art or as described herein. The nomenclatures used in connection with, and the laboratory procedures and techniques of, analytical chemistry, synthetic organic chemistry, and medicinal and pharmaceutical chemistry described herein are those well known and commonly used in the art. Standard techniques are used for chemical syntheses, chemical analyses, pharmaceutical preparation, formulation, and delivery, and treatment of patients.
The term “human TNF-α” (abbreviated herein as hTNF-α) includes a trimeric cytokine protein. The term includes a homotrimeric protein comprising three 17.5 kD TNF-α proteins. The homotrimeric protein is referred to as a “TNF-α protein”. The term human “TNF-α” is intended to include recombinant human TNF-α (rhTNF-α), which can be prepared by standard recombinant expression methods. The sequence of human TNF-α is shown in Table 1.
TABLE 1
Sequence of Human TNF-α
Pro-
Sequence
Sequence
tein
Identifier
12345678901234567890123456789012
Human
SEQ ID
VRSSSRTPSDKPVAHVVANPQAEGQLQWLNDR
TNF-α
NO.: 1
ANALLANGVELRDNQLVVPSEGLYLIYSQVLF
KGQGCPSTHVLLTHTISRIAVSYQTKVNLLSA
IKSPCQRETPEGAEAKPWYEPIYLGGVFQLEK
GDRLSAEINRPDYLDFAESGQVYFGIIAL
The term “antibody”, broadly refers to any immunoglobulin (Ig) molecule, or antigen binding portion thereof, comprised of four polypeptide chains, two heavy (H) chains and two light (L) chains, or any functional fragment, mutant, variant, or derivation thereof, which retains the essential epitope binding features of an Ig molecule. Such mutant, variant, or derivative antibody formats are known in the art.
In a full-length antibody, each heavy chain is comprised of a heavy chain variable region (abbreviated herein as HCVR or VH) and a heavy chain constant region. The heavy chain constant region is comprised of three domains, CH1, CH2 and CH3. Each light chain is comprised of a light chain variable region (abbreviated herein as LCVR or VL) and a light chain constant region. The light chain constant region is comprised of one domain, CL. The VH and VL regions can be further subdivided into regions of hypervariability, termed complementarity determining regions (CDR), interspersed with regions that are more conserved, termed framework regions (FR). Each VH and VL is composed of three CDRs and four FRs, arranged from amino-terminus to carboxy-terminus in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4. Immunoglobulin molecules can be of any type (e.g., IgG, IgE, IgM, IgD, IgA and IgY), class (e.g., IgG 1, IgG2, IgG 3, IgG4, IgA1 and IgA2) or subclass.
The term “antigen-binding portion” or “antigen-binding region” of a binding protein (or simply “binding protein portion”), refers to one or more fragments of a binding protein that retain the ability to specifically bind to an antigen (e.g., hTNF-α). The antigen-binding function of a binding protein can be performed by fragments of a full-length binding protein. Such binding protein embodiments may also have bispecific, dual specific, or multi-specific formats; specifically binding to two or more different antigens. Examples of binding fragments encompassed within the term “antigen-binding portion” of a binding protein include (i) a Fab fragment, a monovalent fragment consisting of the VL, VH, CL and CH1 domains; (ii) a F(ab′)2 fragment, a bivalent fragment comprising two Fab fragments linked by a disulfide bridge at the hinge region; (iii) a Fd fragment consisting of the VH and CH1 domains; (iv) a Fv fragment consisting of the VL and VH domains of a single arm of an antibody, (v) a dAb fragment (Ward et al. (1989) Nature 341:544-546, Winter et al., PCT publication WO 90/05144 A1), which comprises a single variable domain; and (vi) an isolated complementarity determining region (CDR). Furthermore, although the two domains of the Fv fragment, VL and VH, are coded for by separate genes, they can be joined, using recombinant methods, by a synthetic linker that enables them to be made as a single protein chain in which the VL and VH regions pair to form monovalent molecules (known as single chain Fv (scFv); see, e.g., Bird et al. (1988) Science 242:423-426; and Huston et al. (1988) Proc. Natl. Acad. Sci. USA 85:5879-5883). Such single chain binding proteins are also intended to be encompassed within the term “antigen-binding portion” of a binding protein. Other forms of single chain binding proteins, such as diabodies are also encompassed. Diabodies are bivalent, bispecific binding proteins in which VH and VL domains are expressed on a single polypeptide chain, but using a linker that is too short to allow for pairing between the two domains on the same chain, thereby forcing the domains to pair with complementary domains of another chain and creating two antigen binding sites (see, e.g., Holliger, et al. (1993) Proc. Natl. Acad. Sci. USA 90:6444-6448; Poljak, et al. (1994) Structure 2:1121-1123).
The term “binding protein” refers to a polypeptide comprising one or more antigen-binding portions disclosed herein optionally linked to a linker polypeptide or a constant domain. Linker polypeptides comprise two or more amino acid residues joined by peptide bonds and are used to link one or more antigen binding portions. Such linker polypeptides are well known in the art (see e.g., Holliger, et al. (1993) Proc. Natl. Acad. Sci. USA 90:6444-6448; Poljak, et al. (1994) Structure 2:1121-1123). A constant domain refers to a heavy or light chain constant domain. Human IgG heavy chain and light chain constant domain amino acid sequences are known in the art and represented in Table 2.
TABLE 2
Sequence of Human IgG Heavy Chain Constant
Domain and Light Chain Constant Domain
Sequence
Pro-
Identi-
Sequence
tein
fier
12345678901234567890123456789012
Ig
SEQ ID
ASTKGPSVFFLAPSSKSTSGGTAALGCLVKDY
gamma-1
NO.: 2
FPEPVTVSWNSGALTSGVHTFPAVLQSSGLYS
constant
LSSVVTVPSSSLGTQTYICNVNHKPSNTKVDK
region
KVEPKSCDKTHTCPPCPAPELLGGPSVFLFPP
KPKDTLMISRTPEVTCVVVDVSHEDPEVKFNW
YVDGVEVHNAKTKPREEQYNSTYRVVSVLTVL
HQDWLNGKEYKCKVSNKALPAPIEKTISKAKG
QPREPQVYTLPPSREEMTKNQVSLTCLVKGFY
PSDIAVEWESNGQPENNYKTTPPVLDSDGSFF
LYSKLTVDKSRWQQGNVFSCSVMHEALHNHYT
QKSLSLSPGK
Ig
SEQ ID
ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDY
gamma-1
NO.: 3
FPEPVTVSWNSGALTSGVHTFPAVLQSSGLYS
constant
LSSVVTVPSSSLGTQTYICNVNHKPSNTKVDK
region
KVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPP
mutant
KPKDTLMISRTPEVTCVVVDVSHEDPEVKFNW
YVDGVEVHNAKTKPREEQYNSTYRVVSVLTVL
HQDWLNGKEYKCKVSNKALPAPIEKTISKAKG
QPREPQVYTLPPSREEMTKNQVSLTCLVKGFY
PSDIAVEWESNGQPENNYKTTPPVLDSDGSFF
LYSKLTVDKSRWQQGNVFSCSVMHEALHNHYT
QKSLSLSPGK
Ig Kappa
SEQ ID
TVAAPSVFIFPPSDEQLKSGTASVVCLLNNFY
constant
NO.: 4
PREAKVQWKVDNALQSGNSQESVTEQDSKDST
region
YSLSSTLTLSKADYEKHKVYACEVTHQGLSSP
VTKSFNRGEC
Ig
SEQ ID
QPKAAPSVTLFPPSSEELQANKATLVCLISDF
Lambda
NO.: 5
YPGAVTVAWKADSSPVKAGVETTTPSKQSNNK
constant
YAASSYLSLTPEQWKSHRSYSCQVTHEGSTVE
region
KTVAPTECS
A binding protein, or antigen-binding portion thereof, may be part of a larger immunoadhesion molecule, formed by covalent or noncovalent association of the binding protein or binding protein portion with one or more other proteins or peptides. Examples of such immunoadhesion molecules include use of the streptavidin core region to make a tetrameric scFv molecule (Kipriyanov, et al. (1995) Hum. Antibod. Hybridomas 6:93-101) and use of a cysteine residue, a marker peptide and a C-terminal polyhistidine tag to make bivalent and biotinylated scFv molecules (Kipriyanov, et al. (1994) Mol. Immunol. 31:1047-1058). Antibody portions, such as Fab and F(ab′)2 fragments, can be prepared from whole antibodies using conventional techniques, such as papain or pepsin digestion, respectively, of whole antibodies. Moreover, binding proteins, binding protein portions and immunoadhesion molecules can be obtained using standard recombinant DNA techniques, as described herein.
An “isolated binding protein” refers to a binding protein, or antigen-binding portion thereof, that is substantially free of other binding proteins having different antigenic specificities (e.g., an isolated binding protein that specifically binds hTNF-α is substantially free of binding proteins that specifically bind antigens other than hTNF-α). An isolated binding protein that specifically binds hTNF-α may, however, have cross-reactivity to other antigens, such as TNF-α molecules from other species. Moreover, an isolated binding protein may be substantially free of other cellular material and/or chemicals.
The term “human binding protein” includes binding proteins, or antigen-binding portion thereof, that having variable and constant regions derived from human germline immunoglobulin sequences. The human binding proteins disclosed herein may include amino acid residues not encoded by human germline immunoglobulin sequences (e.g., mutations introduced by random or site-specific mutagenesis in vitro or by somatic mutation in vivo), for example in the CDRs and in particular CDR3. However, the term “human binding protein”, is not intended to include binding proteins in which CDR sequences derived from the germline of another mammalian species, such as a mouse, have been grafted onto human framework sequences.
The terms “Kabat numbering”, “Kabat definitions” and “Kabat labeling” are used interchangeably herein. These terms, which are recognized in the art, refer to a system of numbering amino acid residues which are more variable (i.e., hypervariable) than other amino acid residues in the heavy and light chain variable regions of an antibody, or an antigen binding portion thereof (Kabat et al. (1971) Ann. NY Acad. Sci. 190:382-391 and Kabat, et al. (1991) Sequences of Proteins of Immunological Interest, Fifth Edition, U.S. Department of Health and Human Services, NIH Publication No. 91-3242). See also, Martin, “Protein Sequence and Structure Analysis of Antibody Variable Domains,” In Kontermann and Dübel, eds., Antibody Engineering (Springer-Verlag, Berlin, 2001), Chapter 31, especially pages 432-433. For the heavy chain variable region, the hypervariable region ranges from amino acid positions 31 to 35 for CDR1, amino acid positions 50 to 65 for CDR2, and amino acid positions 95 to 106 for CDR3. For the light chain variable region, the hypervariable region ranges from amino acid positions 24 to 34 for CDR1, amino acid positions 50 to 56 for CDR2, and amino acid positions 89 to 97 for CDR3.
The term “CDR” refers to the complementarity determining region within antibody variable sequences. There are three CDRs in each of the variable regions of the heavy chain and the light chain, which are designated CDR1, CDR2 and CDR3, for each of the variable regions. The term “CDR set” refers to a group of three CDRs that occur in a single variable region capable of binding the antigen. The exact boundaries of these CDRs have been defined differently according to different systems. The system described by Kabat (Kabat et al., Sequences of Proteins of Immunological Interest (National Institutes of Health, Bethesda, Md. (1987) and (1991)) not only provides an unambiguous residue numbering system applicable to any variable region of an antibody, but also provides precise residue boundaries defining the three CDRs. These CDRs may be referred to as Kabat CDRs. Chothia and coworkers (Chothia and Lesk (1987) J. Mol. Biol. 196:901-917) and Chothia et al. (1989) Nature 342:877-883) found that certain sub-portions within Kabat CDRs adopt nearly identical peptide backbone conformations, despite having great diversity at the level of amino acid sequence. These sub-portions were designated as L1, L2 and L3 or H1, H2 and H3 where the “L” and the “H” designates the light chain and the heavy chains regions, respectively. These regions may be referred to as Chothia CDRs, which have boundaries that overlap with Kabat CDRs. Other boundaries defining CDRs overlapping with the Kabat CDRs have been described by Padlan (1995) FASEB J. 9:133-139 and MacCallum (1996) J. Mol. Biol. 262(5):732-745. Still other CDR boundary definitions may not strictly follow one of the above systems, but will nonetheless overlap with the Kabat CDRs, although they may be shortened or lengthened in light of prediction or experimental findings that particular residues or groups of residues or even entire CDRs do not significantly impact antigen binding. The methods used herein may utilize CDRs defined according to any of these systems, although particular embodiments use Kabat or Chothia defined CDRs.
Human heavy chain and light chain acceptor sequences are known in the art. In one embodiment of the disclosure the human heavy chain and light chain acceptor sequences are selected from the sequences listed from V-base (hvbase.mrc-cpe.cam.ac.uk/) or from IMGT®, the international ImMunoGeneTics information System® (himgt.cines.fr/textes/IMGTrepertoire/LocusGenes/). In another embodiment of the disclosure the human heavy chain and light chain acceptor sequences are selected from the sequences described in Table 3 and Table 4, respectively.
TABLE 3
Heavy Chain Acceptor Sequences
SEQ
Protein
Sequence
ID No.
region
12345678901234567890123456789012
SEQ ID
VH4-59 FR1
QVQLQESGPGLVKPSETLSLTCTVSGGSISS
NO: 6
SEQ ID
VH4-59 FR2
WIRQPPGKGLEWIG
NO: 7
SEQ ID
VH4-59 FR3
RVTISVDTSKNQFSLKLSSVTAADTAVYYCAR
NO: 8
SEQ ID
VH3-53 FR1
EVQLVESGGGLIQPGGSLRLSCAASGFTVSS
NO: 9
SEQ ID
VH3-53 FR2
WVRQAPGKGLEWVS
NO: 10
SEQ ID
VH3-53 FR3
RFTISRDNSKNTLYLQMNSLRAEDTAVYYCAR
NO: 11
SEQ ID
JH1/JH4/
WGQGTLVTVSS
NO: 12
JH5 FR4
SEQ ID
JH2 FR4
WGRGTLVTVSS
NO: 13
SEQ ID
JH6 FR4
WGQGTTVTVSS
NO: 14
TABLE 4
Light Chain Acceptor Sequences
SEQ
Protein
Sequence
ID No.
region
12345678901234567890123456789012
SEQ ID
1-39/O12
DIQMTQSPSSLSASVGDRVTITC
NO: 15
FR1
SEQ ID
1-39/O12
WYQQKPGKAPKLLIY
NO: 16
FR2
SEQ ID
1-39/O12
GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC
NO: 17
FR3
SEQ ID
3-15/L2 FR1
EIVMTQSPATLSVSPGERATLSC
NO: 18
SEQ ID
3-15/L2 FR2
WYQQKPGQAPRLLIY
NO: 19
SEQ ID
3-15/L2 FR3
GIPARFSGSGSGTEFTLTISSLQSEDFAVYYC
NO: 20
SEQ ID
JK2 FR4
FGQGTKLEIKR
NO: 21
The term “multivalent binding protein” is used in this specification to denote a binding protein comprising two or more antigen binding sites. The multivalent binding protein may be engineered to have the three or more antigen binding sites, and is generally not a naturally occurring antibody. The term “multispecific binding protein” refers to a binding protein capable of binding two or more related or unrelated targets. Dual variable domain (DVD) binding proteins or immunoglobulins (DVD-Ig) as used herein, are binding proteins that comprise two or more antigen binding sites and are tetravalent or multivalent binding proteins. Such DVD-binding proteins may be monospecific, i.e., capable of binding one antigen or multispecific, i.e., capable of binding two or more antigens. DVD-binding proteins comprising two heavy chain DVD-Ig polypeptides and two light chain DVD-Ig polypeptides are referred to a DVD-Ig. Each half of a DVD-Ig comprises a heavy chain DVD-Ig polypeptide, and a light chain DVD-Ig polypeptide, and two antigen binding sites. Each binding site comprises a heavy chain variable domain and a light chain variable domain with a total of 6 CDRs involved in antigen binding per antigen binding site. DVD binding proteins and methods of making DVD binding proteins are disclosed in U.S. Pat. No. 7,612,181.
One aspect of the disclosure pertains to a DVD binding protein comprising binding proteins capable of binding TNF-α. In a particular embodiment, the DVD binding protein is capable of binding TNF-α and a second target.
The term “neutralizing” refers to neutralization of a biological activity of a cytokine when a binding protein specifically binds the cytokine. In a particular embodiment, binding of a neutralizing binding protein to hTNF-α results in inhibition of a biological activity of hTNF-α, e.g., the neutralizing binding protein binds hTNF-α and reduces a biologically activity of hTNF-α by at least about 20%, 40%, 60%, 80%, 85% or more Inhibition of a biological activity of hTNF-α by a neutralizing binding protein can be assessed by measuring one or more indicators of hTNF-α biological activity well known in the art. For example neutralization of the cytoxicity of TNF-α on L929 cells.
In another embodiment, the terms “agonist” or “agonizing” refer to an increase of a biological activity of TNF-α when a binding protein specifically binds TNF-α, e.g., hTNF-α. In a particular embodiment, binding of an agonizing binding protein to TNF-α results in the increase of a biological activity of TNF-α. In a particular embodiment, the agonistic binding protein binds TNF-α and increases a biologically activity of TNF-α by at least about 20%, 40%, 60%, 80%, 85%, 90%, 95, 96%, 97%, 98%, 99%, and 100%. An inhibition of a biological activity of TNF-α by an agonistic binding protein can be assessed by measuring one or more indicators of TNF-α biological activity well known in the art.
The term “activity” includes activities such as the binding specificity/affinity of a binding protein for an antigen, for example, a hTNF-α binding protein that binds to a TNF-α antigen and/or the neutralizing potency (or agonizing potency) of a binding protein, for example, a hTNF-α binding protein whose binding to hTNF-α inhibits the biological activity of hTNF-α, e.g., neutralization of the cytoxicity of TNF-α on L929 cells.
The term “surface plasmon resonance” refers to an optical phenomenon that allows for the analysis of real-time bio specific interactions by detection of alterations in protein concentrations within a biosensor matrix, for example using the BIAcore system (Pharmacia Biosensor AB, Uppsala, Sweden and Piscataway, N.J.).
The term “Kon” refers to the on rate constant for association of a binding protein (e.g., an antibody) to the antigen to form, e.g., the antibody/antigen complex as is known in the art. The “Kon” also is known by the terms “association rate constant”, or “ka”, as used interchangeably herein. This value indicating the binding rate of an antibody to its target antigen or the rate of complex formation between an antibody and antigen also is shown by the equation below:
Antibody(“Ab”)+Antigen(“Ag”)→Ab−Ag
The term “Koff” refers to the off rate constant for dissociation, or “dissociation rate constant”, of a binding protein (e.g., an antibody), from the, e.g., antibody/antigen complex as is known in the art. This value indicates the dissociation rate of an antibody from its target antigen or separation of Ab−Ag complex over time into free antibody and antigen as shown by the equation below:
Ab+Ag←Ab−Ag
The term “KD” refers to the “equilibrium dissociation constant” and refers to the value obtained in a titration measurement at equilibrium, or by dividing the dissociation rate constant (Koff) by the association rate constant (Kon). The association rate constant, the dissociation rate constant and the equilibrium dissociation constant are used to represent the binding affinity of an antibody to an antigen. Methods for determining association and dissociation rate constants are well known in the art. Using fluorescence-based techniques offers high sensitivity and the ability to examine samples in physiological buffers at equilibrium. Other experimental approaches and instruments such as a BIAcore® (biomolecular interaction analysis) assay can be used (e.g., instrument available from BIAcore International AB, a GE Healthcare company, Uppsala, Sweden). Additionally, a KinExA® (Kinetic Exclusion Assay) assay, available from Sapidyne Instruments (Boise, Id.) can also be used.
I. Binding Proteins that Bind Human TNF-α
One aspect of the present disclosure provides isolated fully-human anti-human TNF binding proteins, such as monoclonal antibodies, or antigen-binding portions thereof, that bind to TNF-α with high affinity, a slow off rate and high neutralizing capacity. A second aspect of the disclosure provides affinity-matured fully-human anti-TNF binding proteins, such as monoclonal antibodies, or antigen-binding portions thereof, that bind to TNF-α with high affinity, a slow off rate and high neutralizing capacity.
A. Method of Making TNF-α Binding Proteins
The binding proteins disclosed herein may be made by any of a number of techniques known in the art.
1. Anti-TNF-α Monoclonal Antibodies Using Transgenic Animals
In another embodiment of the disclosure, binding proteins are produced by immunizing a non-human animal comprising some, or all, of the human immunoglobulin locus with a TNF-α antigen. In a particular embodiment, the non-human animal is a XENOMOUSE transgenic mouse, an engineered mouse strain that comprises large fragments of the human immunoglobulin loci and is deficient in mouse antibody production. See, e.g., Green et al. (1994) Nature Genet. 7:13-21 and U.S. Pat. Nos. 5,916,771; 5,939,598; 5,985,615; 5,998,209; 6,075,181; 6,091,001; 6,114,598 and 6,130,364. See also PCT Publications WO 91/10741, published Jul. 25, 1991; WO 94/02602, published Feb. 3, 1994; WO 96/34096 and WO 96/33735, both published Oct. 31, 1996; WO 98/16654, published Apr. 23, 1998; WO 98/24893, published Jun. 11, 1998; WO 98/50433, published Nov. 12, 1998; WO 99/45031, published Sep. 10, 1999; WO 99/53049, published Oct. 21, 1999; WO 00/09560, published Feb. 24, 2000; and WO 00/37504, published Jun. 29, 2000. The XENOMOUSE transgenic mouse produces an adult-like human repertoire of fully human antibodies, and generates antigen-specific human Mabs. The XENOMOUSE transgenic mouse contains approximately 80% of the human antibody repertoire through introduction of megabase sized, germline configuration YAC fragments of the human heavy chain loci and x light chain loci. See, Mendez et al. (1997) Nature Genet. 15:146-156; Green and Jakobovits (1998) J. Exp. Med. 188:483-495.
2. Anti-TNF-α Monoclonal Antibodies Using Recombinant Antibody Libraries
In vitro methods also can be used to make the binding protein disclosed herein, wherein an antibody library is screened to identify an antibody having the desired binding specificity. Methods for such screening of recombinant antibody libraries are well known in the art and include methods described in, for example, U.S. Pat. No. 5,223,409; PCT Publications WO 92/18619; WO 91/17271; WO 92/20791; WO 92/15679; WO 93/01288; WO 92/01047; WO 92/09690; and WO 97/29131; Fuchs et al. (1991) Bio/Technology 9:1369-1372; Hay et al. (1992) Hum. Antibod. Hybridomas 3:81-85; Huse et al. (1989) Science 246:1275-1281; McCafferty et al. (1990) Nature 348:552-554; Griffiths et al. (1993) EMBO J. 12:725-734; Hawkins et al. (1992) J. Mol. Biol. 226:889-896; Clackson et al. (1991) Nature 352:624-628; Gram et al. (1992) Proc. Natl. Acad. Sci. USA 89:3576-3580; Garrard et al. (1991) Bio/Technology 9:1373-1377; Hoogenboom et al. (1991) Nucl. Acid Res. 19:4133-4137; and Barbas et al. (1991) Proc. Natl. Acad. Sci. USA 88:7978-7982; and U.S. Patent Publication No. 2003.0186374.
The recombinant antibody library may be from a subject immunized with TNF-α, or a portion of TNF-α. Alternatively, the recombinant antibody library may be from a naïve subject, i.e., one who has not been immunized with TNF-α, such as a human antibody library from a human subject who has not been immunized with human TNF-α. Antibodies disclosed herein are selected by screening the recombinant antibody library with the peptide comprising human TNF-α to thereby select those antibodies that recognize TNF-α. Methods for conducting such screening and selection are well known in the art, such as described in the references in the preceding paragraph. To select antibodies disclosed herein having particular binding affinities for hTNF-α, such as those that dissociate from human TNF-α with a particular koff rate constant, the art-known method of surface plasmon resonance can be used to select antibodies having the desired koff rate constant. To select antibodies disclosed herein having a particular neutralizing activity for hTNF-α, such as those with a particular an IC50, standard methods known in the art for assessing the inhibition of hTNF-α activity may be used.
In one aspect, provided is an isolated binding protein, or an antigen-binding portion thereof, that binds TNF-α, e.g., human TNF-α. In a particular embodiment, the binding protein is a neutralizing binding protein. In various embodiments, the binding protein is a recombinant binding protein or a monoclonal antibody.
For example, the binding proteins disclosed herein can also be generated using various phage display methods known in the art. In phage display methods, functional antibody domains are displayed on the surface of phage particles which carry the polynucleotide sequences encoding them. In a particular, such phage can be utilized to display antigen-binding domains expressed from a repertoire or combinatorial antibody library (e.g., human or murine). Phage expressing an antigen binding domain that binds the antigen of interest can be selected or identified with antigen, e.g., using labeled antigen or antigen bound or captured to a solid surface or bead. Phage used in these methods are typically filamentous phage including fd and M13 binding domains expressed from phage with Fab, Fv or disulfide stabilized Fv antibody domains recombinantly fused to either the phage gene III or gene VIII protein. Examples of phage display methods that can be used to make the binding proteins disclosed herein can be found in the art.
As described in the above references, after phage selection, the binding protein coding regions from the phage can be isolated and used to generate whole binding proteins including human binding protein or any other desired antigen binding fragment, and expressed in any desired host, including mammalian cells, insect cells, plant cells, yeast, and bacteria, e.g., as described in detail below. For example, techniques to recombinantly produce Fab, Fab′ and F(ab′)2 fragments can also be employed using methods known in the art such as those disclosed in PCT Publication WO 92/22324; Mullinax et al. (1992) BioTechniques 12(6):864-869; and Sawai et al. (1995) Am. J. Reprod. Immunol. 34:26-34; and Better et al. (1998) Science 240:1041-1043. Examples of techniques which can be used to produce single-chain Fvs and antibodies include those described in U.S. Pat. Nos. 4,946,778 and 5,258,498; Huston et al. (1991) Methods Enzymol. 203:46-88; Shu et al. (1993) Proc. Natl. Acad Sci. USA 90:7995-7999; and Skerra et al. (1998) Science 240:1038-1041.
Alternative to screening of recombinant antibody libraries by phage display, other methodologies known in the art for screening large combinatorial libraries can be applied to the identification of dual specificity binding protein disclosed herein. One type of alternative expression system is one in which the recombinant antibody library is expressed as RNA-protein fusions, as described in PCT Publication No. WO 98/31700 and in Roberts and Szostak (1997) Proc. Natl. Acad. Sci. USA 94:12297-12302. In this system, a covalent fusion is created between an mRNA and the peptide or protein that it encodes by in vitro translation of synthetic mRNAs that carry puromycin, a peptidyl acceptor antibiotic, at their 3′ end. Thus, a specific mRNA can be enriched from a complex mixture of mRNAs (e.g., a combinatorial library) based on the properties of the encoded peptide or protein, e.g., antibody, or portion thereof, such as binding of the antibody, or portion thereof, to the dual specificity antigen. Nucleic acid sequences encoding antibodies, or portions thereof, recovered from screening of such libraries can be expressed by recombinant means as described above (e.g., in mammalian host cells) and, moreover, can be subjected to further affinity maturation by either additional rounds of screening of mRNA-peptide fusions in which mutations have been introduced into the originally selected sequence(s), or by other methods for affinity maturation in vitro of recombinant antibodies, as described above.
In another approach the binding proteins disclosed herein can also be generated using yeast display methods known in the art. In yeast display methods, genetic methods are used to tether antibody domains to the yeast cell wall and display them on the surface of yeast. In particular, such yeast can be utilized to display antigen-binding domains expressed from a repertoire or combinatorial antibody library (e.g., human or murine). Examples of yeast display methods that can be used to make the binding proteins disclosed herein include those disclosed Wittrup et al. U.S. Pat. No. 6,699,658 and Frenken et al., U.S. Pat. No. 6,114,147.
B. Production of Recombinant TNF-α Binding Proteins
Binding proteins disclosed herein may be produced by any of a number of techniques known in the art. For example, expression from host cells, wherein expression vector(s) encoding the heavy and light chains is (are) transfected into a host cell by standard techniques. The various forms of the term “transfection” are intended to encompass a wide variety of techniques commonly used for the introduction of exogenous DNA into a prokaryotic or eukaryotic host cell, e.g., electroporation, calcium-phosphate precipitation, DEAE-dextran transfection and the like. Although it is possible to express the binding proteins disclosed herein in either prokaryotic or eukaryotic host cells, expression of binding protein in eukaryotic cells is contemplated, for example, in mammalian host cells, because such eukaryotic cells (and in particular mammalian cells) are more likely than prokaryotic cells to assemble and secrete a properly folded and immunologically active binding protein.
Mammalian host cells for expressing the recombinant binding proteins disclosed herein include Chinese Hamster Ovary (CHO cells) (including dhfr-CHO cells, described in Urlaub and Chasin (1980) Proc. Natl. Acad. Sci. USA 77:4216-4220, used with a DHFR selectable marker, e.g., as described in Kaufman and Sharp (1982) J. Mol. Biol. 159:601-621), NS0 myeloma cells, COS cells and SP2 cells. When recombinant expression vectors encoding binding protein genes are introduced into mammalian host cells, the binding proteins are produced by culturing the host cells for a period of time sufficient to allow for expression of the binding protein in the host cells or, in particular, secretion of the antibody into the culture medium in which the host cells are grown. Antibodies can be recovered from the culture medium using standard protein purification methods.
Host cells can also be used to produce functional binding protein fragments, such as Fab fragments or scFv molecules. It will be understood that variations on the above procedure are within the scope of the present disclosure. For example, it may be desirable to transfect a host cell with DNA encoding functional fragments of either the light chain and/or the heavy chain of a binding protein disclosed herein. Recombinant DNA technology may also be used to remove some, or all, of the DNA encoding either or both of the light and heavy chains that is not necessary for binding to the antigens of interest. The molecules expressed from such truncated DNA molecules are also encompassed by the binding proteins disclosed herein. In addition, bifunctional binding proteins may be produced in which one heavy and one light chain are a binding protein disclosed herein and the other heavy and light chain are specific for an antigen other than the antigens of interest by crosslinking a binding protein disclosed herein to a second binding protein by standard chemical crosslinking methods.
In an exemplary system for recombinant expression of a binding protein, or antigen-binding portion thereof, disclosed herein, a recombinant expression vector encoding both the heavy chain and the light chain is introduced into dhfr CHO cells by calcium phosphate-mediated transfection. Within the recombinant expression vector, the heavy and light chain genes are each operatively linked to CMV enhancer/AdMLP promoter regulatory elements to drive high levels of transcription of the genes. The recombinant expression vector also carries a DHFR gene, which allows for selection of CHO cells that have been transfected with the vector using methotrexate selection/amplification. The selected transformant host cells are cultured to allow for expression of the heavy and light chains and intact binding protein is recovered from the culture medium. Standard molecular biology techniques are used to prepare the recombinant expression vector, transfect the host cells, select for transformants, culture the host cells and recover the binding protein from the culture medium. Still further a method of synthesizing a recombinant binding protein disclosed herein is provided by culturing a host cell disclosed herein in a suitable culture medium until a recombinant binding protein disclosed herein is synthesized. The method can further comprise isolating the recombinant binding protein from the culture medium.
II. hTNF-α Binding Proteins
A. Individual Clone Sequences
Table 5 provides the VH and VL sequences of fully human anti-human TNF binding proteins, including CDRs from each VH and VL sequence.
TABLE 5
Individual Fully Human Anti-TNF-α VH Sequences
Sequence
Protein region
123456789012345678901234567890
AE11-1 VH
SEQ ID NO.: 22
EVQLVQSGAEVKKPGASVKVSCKASGYTFT
SYDVNWVRQATGQGLEWMGWMNPNSGNTGY
AQKFQGRVTITADESTSTAYMELSSLRSED
TAVYYCAIFDSDYMDVWGKGTLVTVSS
AE11-1 VH
CDR-
Residues 31-35
SYDVN
H1
of SEQ ID
NO.: 22
AE11-1 VH
CDR-
Residues 50-66
WMNPNSGNTGYAQKFQG
H2
of SEQ ID
NO.: 22
AE11-1 VH
CDR-
Residues 99-106
FDSDYMDV
H3
of SEQ ID
NO.: 22
AE11-1 VL
SEQ ID NO.: 23
SYELTQPPSVSLSPGQTARITCSGDALPKQ
YAYWYQQKPGQAPVLVIYKDTERPSGIPER
FSGSSSGTTVTLTISGAQAEDEADYYCQSA
DSSGTSWVFGGGTKLTVL
AE11-1 VL
CDR-
Residues 23-33
SGDALPKQYAY
L1
of SEQ ID
NO.: 23
AE11-1 VL
CDR-
Residues 49-55
KDTERPS
L2
of SEQ ID
NO.: 23
AE11-1 VL
CDR-
Residues 89-98
SADSSGTSWV
L3
of SEQ ID
NO.: 23
AE11-5 VH
SEQ ID NO.: 24
EVQLVQSGAEVKKPGSSAKVSCKASGGTFS
SYAISWVRQAPGQGLEWMGGIIPILGTANY
AQKFLGRVTITADESTSTVYMELSSLRSED
TAVYYCARGLYYDPTRADYWGQGTLVTVSS
AE11-5 VH
CDR-
Residues 31-35
SYAIS
H1
of SEQ ID
NO.: 24
AE11-5 VH
CDR-
Residues 50-66
GIIPILGTANYAQKFLG
H2
of SEQ ID
NO.: 24
AE11-5 VH
CDR-
Residues 99-109
GLYYDPTRADY
H3
of SEQ ID
NO.: 24
AE11-5 VL
SEQ ID NO.: 25
DIVMTQSPDFHSVTPKEKVTITCRASQSIG
SSLHWYQQKPDQSPKLLIRHASQSISGVPS
RFSGSGSGTDFTLTIHSLEAEDAATYYCHQ
SSSSPPPTFGQGTQVEIK
AE11-5 VL
CDR-
Residues 24-34
RASQSIGSSLH
L1
of SEQ ID
NO.: 25
AE11-5 VL
CDR-
Residues 50-56
HASQSIS
L2
of SEQ ID
NO.: 25
AE11-5 VL
CDR-
Residues 89-98
HQSSSSPPPT
L3
of SEQ ID
NO.: 25
TNF-JK1 VH
SEQ ID NO.: 26
EVQLVESGGGLVQPGGSLRLSCATSGFTFN
NYWMSWVRQAPGKGLEWVANINHDESEKYY
VDSAKGRFTISRDNAEKSLFLQMNSLRAED
TAVYYCARIIRGRVGFDYYNYAMDVWGQGT
LVTVSS
TNF-JK1 VH
CDR-
Residues 31-35
NYWMS
H1
of SEQ ID
NO.: 26
TNF-JK1 VH
CDR-
Residues 50-66
NINHDESEKYYVDSAKG
H2
of SEQ ID
NO.: 26
TNF-JK1 VH
CDR-
Residues 99-115
IIRGRVGFDYYNYAMDV
H3
of SEQ ID
NO.: 26
TNF-JK1 VL
SEQ ID NO.: 27
DIRLTQSPSPLSASVGDRVTITCRASQSIG
NYLNWYQHKPGKAPKLLIYAASSLQSGVPS
RFSGTGSGTDFTLTISSLQPEDFATYYCQE
SYSLIFAGGTKVEIK
TNF-JK1 VL
CDR-
Residues 24-34
RASQSIGNYLN
L1
of SEQ ID
NO.: 27
TNF-JK1 VL
CDR-
Residues 50-56
AASSLQS
L2
of SEQ ID
NO.: 27
TNF-JK1 VL
CDR-
Residues 89-95
QESYSLI
L3
of SEQ ID
NO.: 27
TNF-Y7C VH
SEQ ID NO.: 28
EVQLVQSGAEVKKPGASVKVSCKTSGYTFS
NYDINWVRQPTGQGLEWMGWMDPNNGNTGY
AQKFVGRVTMTRDTSKTTAYLELSGLKSED
TAVYYCARSSGSGGTWYKEYFQSWGQGTMV
TVSS
TNF-Y7C VH
CDR-
Residues 31-35
NYDIN
H1
of SEQ ID
NO.: 28
TNF-Y7C VH
CDR-
Residues 50-66
WMDPNNGNTGYAQKFVG
H2
of SEQ ID
NO.: 28
TNF-Y7C VH
CDR-
Residues 99-112
KSSGSGGTWYKEYFQS
H3
of SEQ ID
NO.: 28
TNF-Y7C VL
SEQ ID NO.: 29
DIVMTQSPLSLPVTPGEPASISCRSSQSLL
HSNGYNYLDWYLQKPGQFPQLLIYLGSYRA
SGVPDRFSGSGSGTDFTLKISRVEAEDVGV
YYCMQRIEFPPGTFGQGTKLGIK
TNF-Y7C VL
CDR-
Residues 24-39
RSSQSLLHSNGYNYLD
L1
of SEQ ID
NO.: 29
TNF-Y7C VL
CDR-
Residues 55-61
LGSYRAS
L2
of SEQ ID
NO.: 29
TNF-Y7C VL
CDR-
Residues 94-103
MQRIEFPPGT
L3
of SEQ ID
NO.: 29
AE11-7 VH
SEQ ID NO.: 30
EVQLVQSGAEVKKPGASVKVSCKTSGYSLT
QYPIHWVRQAPGQRPEWMGWISPGNGNTKL
SPKFQGRVTLSRDASAGTVFMDLSGLTSDD
TAVYFCTSVDLGDHWGQGTLVTVSS
AE11-7 VH
CDR-
Residues 31-35
QYPIH
H1
of SEQ ID
NO.: 30
AE11-7 VH
CDR-
Residues 50-66
WISPGNGNTKLSPKFQG
H2
of SEQ ID
NO.: 30
AE11-7 VH
CDR-
Residues 99-104
VDLGDH
H3
of SEQ ID
NO.: 30
AE11-7 VL
SEQ ID NO.: 31
DIVMTQSPEFQSVTPKEKVTITCRASQSIG
SSLHWYQQKPDQSPKLLINYASQSFSGVPS
RFSGGGSGTDFTLTINSLEAEDAATYYCHQ
SSNLPITFGQGTRLEIK
AE11-7 VL
CDR-
Residues 24-34
RASQSIGSSLH
L1
of SEQ ID
NO.: 31
AE11-7 VL
CDR-
Residues 50-56
YASQSFS
L2
of SEQ ID
NO.: 31
AE11-7 VL
CDR-
Residues 89-97
HQSSNLPIT
L3
of SEQ ID
NO.: 31
AE11-13 VH
SEQ ID NO.: 32
EVQLVESGGGLVQPGRSLRLSCAASGFTFD
DYPMHWVRQAPGEGLEWVSGISSNSASIGY
ADSVKGRFTISRDNAQNTLYLQMNSLGDED
TAVYYCVSLTLGIGQGTLVTVSS
AE11-13 VH
CDR-
Residues 31-35
DYPMH
H1
of SEQ ID
NO.: 32
AE11-13 VH
CDR-
Residues 50-66
GISSNSASIGYADSVKG
H2
of SEQ ID
NO.: 32
AE11-13 VH
CDR-
Residues 99-102
LTLG
H3
of SEQ ID
NO.: 32
AE11-13 VL
SEQ ID NO.: 33
DIRLTQSPSSLSASVGDRVTITCRASQSIG
NYLHWYQQKPGKAPKLLIYAASSLQSGVPS
RFSGSGSGTDFTLTISSLQPEDFATYYCQQ
SYSTLYSFGQGTKLEIK
AE11-13 VL
CDR-
Residues 24-34
RASQSIGNYLH
L1
of SEQ ID
NO.: 33
AE11-13 VL
CDR-
Residues 50-56
AASSLQS
L2
of SEQ ID
NO.: 33
AE11-13 VL
CDR-
Residues 89-97
QQSYSTLYS
L3
of SEQ ID
NO.: 33
B. IgG Converted Clones
Table 6 provides the VH sequence of humanized anti-TNF MAK-195 antibodies that were converted into IgG clones as discussed in detail in Example 2.
TABLE 6
Humanized anti-TNF MAK-195 Ab VH sequences of
IgG converted clones
Sequence
Protein region
123456789012345678901234567890
A8
SEQ ID NO.: 34
EVQLVESGGGLVQPGGSLRLSCAASGFTFS
VH
NYGVNWVRQAPGKGLEWVSMIAADGFTDYA
SSVKGRFTISRDNSKNTLYLQMNSLRAEDT
AVYYCAREWHHGPVAYWGQGTLVTVSS
A8
CDR-H1
Residues 31-35
NYGVN
VH
of SEQ ID
NO.: 34
A8
CDR-H2
Residues 50-65
MIAADGFTDYASSVKG
VH
of SEQ ID
NO.: 34
A8
CDR-H3
Residues 98-106
EWHHGPVAY
VH
of SEQ ID
NO.: 34
B5
SEQ ID NO.: 35
EVQLVESGGGLVQPGGSLRLSCAASGFTFS
VH
NYGVSWVRQAPGKGLEWVSLIRGDGSTDYA
SSLKGRFTISRDNSKNTLYLQMNSLRAEDT
AVYYCAREWHHGPVAYWGQGTLVTVSS
B5
CDR-H1
Residues 31-35
NYGVS
VH
of SEQ ID
NO.: 35
B5
CDR-H2
Residues 50-65
LIRGDGSTDYASSLKG
VH
of SEQ ID
NO.: 35
B5
CDR-H3
Residues 98-106
EWHHGPVAY
VH
of SEQ ID
NO.: 35
rHC44
SEQ ID NO.: 36
EVQLVESGGGLVQPGGSLRLSCAASGFTFS
VH
NYGVSWVRQAPGKGLEWVSMIWADGSTHYA
DTLKSRFTISRDNSKNTLYLQMNSLRAEDT
AVYYCAREWQHGPVAYWGQGTLVTVSS
rHC44
CDR-H1
Residues 31-35
NYGVS
VH
of SEQ ID
NO.: 36
rHC44
CDR-H2
Residues 50-65
MIWADGSTHYADTLKS
VH
of SEQ ID
NO.: 36
rHC44
CDR-H3
Residues 98-106
EWQHGPVAY
VH
of SEQ ID
NO.: 36
rHC22
SEQ ID NO.: 37
EVQLVESGGGLVQPGGSLRLSCAASGFTFS
VH
NYGVTWVRQAPGKGLEWVSMIWADGSTDYA
DTVKGRFTISRDNSKNTLYLQMNSLRAEDT
AVYYCAREWQHGPVAYWGQGTLVTVSS
rHC22
CDR-H1
Residues 31-35
NYGVT
VH
of SEQ ID
NO.: 37
rHC22
CDR-H2
Residues 50-65
MIWADGSTDYADTVKG
VH
of SEQ ID
NO.: 37
rHC22
CDR-H3
Residues 98-106
EWQHGPVAY
VH
of SEQ ID
NO.: 37
rHC81
SEQ ID NO.: 38
EVQLVESGGGLVQPGGSLRLSCAASGFTFS
VH
NYGVTWVRQAPGKGLEWVSMIWADGSTHYA
DSVKSRFTISRDNSKNTLYLQMNSLRAEDT
AVYYCAREWQHGPLAYWGQGTLVTVSS
rHC81
CDR-H1
Residues 31-35
NYGVT
VH
of SEQ ID
NO.: 38
rHC81
CDR-H2
Residues 50-65
MIWADGSTHYADSVKS
VH
of SEQ ID
NO.: 38
rHC81
CDR-H3
Residues 98-106
EWQHGPLAY
VH
of SEQ ID
NO.: 38
rHC18
SEQ ID NO.: 39
EVQLVESGGGLVQPGGSLRLSCAASGFTFS
VH
NYGVTWVRQAPGKGLEWVSMIWSDGSTDYA
SSVKGRFTISRDNSKNTLYLQMNSLRAEDT
AVYYCAREWQHGPVAYWGQGTLVTVSS
rHC18
CDR-H1
Residues 31-35
NYGVT
VH
of SEQ ID
NO.: 39
rHC18
CDR-H2
Residues 50-65
MIWSDGSTDYASSVKG
VH
of SEQ ID
NO.: 39
rHC18
CDR-H3
Residues 98-106
EWQHGPVAY
VH
of SEQ ID
NO.: 39
rHC14
SEQ ID NO.: 40
EVQLVESGGGLVQPGGSLRLSCAASGFTFS
VH
NYGVTWVRQAPGKGLEWVSMIWADGSTHYA
SSLKGRFTISRDNSKNTLYLQMNSLRAEDT
AVYYCAREWQHGPAAYWGQGTLVTVSS
rHC14
CDR-H1
Residues 31-35
NYGVT
VH
of SEQ ID
NO.: 40
rHC14
CDR-H2
Residues 50-65
MIWADGSTHYASSLKG
VH
of SEQ ID
NO.: 40
rHC14
CDR-H3
Residues 98-106
EWQHGPAAY
VH
of SEQ ID
NO.: 40
rHC3
SEQ ID NO.: 41
EVQLVESGGGLVQPGGSLRLSCAASGFTFS
VH
NYGVSWVRQAPGKGLEWVSMIWADGSTHYA
SSLKGRFTISRDNSKNTLYLQMNSLRAEDT
AVYYCAREWQHGPVAYWGQGTLVTVSS
rHC3
CDR-H1
Residues 31-35
NYGVS
VH
of SEQ ID
NO.: 41
rHC3
CDR-H2
Residues 50-65
MIWADGSTHYASSLKG
VH
of SEQ ID
NO.: 41
rHC3
CDR-H3
Residues 98-106
EWQHGPVAY
VH
of SEQ ID
NO.: 41
rHC19
SEQ ID NO.: 42
EVQLVESGGGLVQPGGSLRLSCAASGFTFS
VH
NYGVTWVRQAPGKGLEWVSMIWADGSTHYA
SSVKGRFTISRDNSKNTLYLQMNSLRAEDT
AVYYCAREWQHGPAAYWGQGTLVTVSS
rHC19
CDR-H1
Residues 31-35
NYGVT
VH
of SEQ ID
NO.: 42
rHC19
CDR-H2
Residues 50-65
MIWADGSTHYASSVKG
VH
of SEQ ID
NO.: 42
rHC19
CDR-H3
Residues 98-106
EWQHGPAAY
VH
of SEQ ID
NO.: 42
rHC34
SEQ ID NO.: 43
EVQLVESGGGLVQPGGSLRLSCAASGFTFS
VH
NYGVTWVRQAPGKGLEWVSMIWADGSTHYA
SSVKGRFTISRDNSKNTLYLQMNSLRAEDT
AVYYCAREWQHGPSAYWGQGTLVTVSS
rHC34
CDR-H1
Residues 31-35
NYGVT
VH
of SEQ ID
NO.: 43
rHC34
CDR-H2
Residues 50-65
MIWADGSTHYASSVKG
VH
of SEQ ID
NO.: 43
rHC34
CDR-H3
Residues 98-106
EWQHGPSAY
VH
of SEQ ID
NO.: 43
rHC83
SEQ ID NO.: 44
EVQLVESGGGLVQPGGSLRLSCAASGFTFS
VH
NYGVTWVRQAPGKGLEWVSMIWADGSTHYA
SSVKGRFTISRDNSKNTLYLQMNSLRAEDT
AVYYCAREWQHGPVAYWGQGTLVTVSS
rHC83
CDR-H1
Residues 31-35
NYGVT
VH
of SEQ ID
NO.: 44
rHC83
CDR-H2
Residues 50-65
MIWADGSTHYASSVKG
VH
of SEQ ID
NO.: 44
rHC83
CDR-H3
Residues 98-106
EWQHGPVAY
VH
of SEQ ID
NO.: 44
S4-19
SEQ ID NO.: 45
EVQLVESGGGLVQPGGSLRLSCAASGFTFS
VH
NYGVEWVRQAPGKGLEWVSGIWADGSTHYA
DTVKSRFTISRDNSKNTLYLQMNSLRAEDT
AVYYCAREWQHGPVAYWGQGTLVTVSS
S4-19
CDR-H1
Residues 31-35
NYGVE
VH
of SEQ ID
NO.: 45
S4-19
CDR-H2
Residues 50-65
GIWADGSTHYADTVKS
VH
of SEQ ID
NO.: 45
S4-19
CDR-H3
Residues 98-106
EWQHGPVAY
VH
of SEQ ID
NO.: 45
S4-50
SEQ ID NO.: 46
EVQLVESGGGLVQPGGSLRLSCAASGFTFS
VH
NYGVEWVRQAPGKGLEWVSGIWADGSTHYA
DTVKSRFTISRDNSKNTLYLQMNSLRAEDT
AVYYCAREWQHGPVGYWGQGTLVTVSS
S4-50
CDR-H1
Residues 31-35
NYGVE
VH
of SEQ ID
NO.: 46
S4-50
CDR-H2
Residues 50-65
GIWADGSTHYADTVKS
VH
of SEQ ID
NO.: 46
S4-50
CDR-H3
Residues 98-106
EWQHGPVGY
VH
of SEQ ID
NO.: 46
S4-63
SEQ ID NO.: 47
EVQLVESGGGLVQPGGSLRLSCAASGFTFS
VH
NYGVEWVRQAPGKGLEWVSGIWADGSTHYA
DTVKSRFTISRDNSKNTLYLQMNSLRAEDT
AVYYCAREWQHGPVGYWGQGTLVTVSS
S4-63
CDR-H1
Residues 31-35
NYGVE
VH
of SEQ ID
NO.: 47
S4-63
CDR-H2
Residues 50-65
GIWADGSTHYADTVKS
VH
of SEQ ID
NO.: 47
S4-63
CDR-H3
Residues 98-106
EWQHGPVGY
VH
of SEQ ID
NO.: 47
S4-55
SEQ ID NO.: 48
EVQLVESGGGLVQPGGSLRLSCAASGFTFS
VH
NYGVTWVRQAPGKGLEWVSMIWADGSTDYA
STVKGRFTISRDNSKNTLYLQMNSLRAEDT
AVYYCAREWQHGPVGYWGQGTLVTVSS
S4-55
CDR-H1
Residues 31-35
NYGVT
VH
of SEQ ID
NO.: 48
S4-55
CDR-H2
Residues 50-65
MIWADGSTDYASTVKG
VH
of SEQ ID
NO.: 48
S4-55
CDR-H3
Residues 98-106
EWQHGPVGY
VH
of SEQ ID
NO.: 48
S4-6
SEQ ID NO.: 49
EVQLVESGGGLVQPGGSLRLSCAASGFTFS
VH
NYGVTWVRQAPGKGLEWVSMIWADGSTHYA
SSVKGRFTISRDNSKNTLYLQMNSLRAEDT
AVYYCAREWQHGPVAYWGQGTLVTVSS
S4-6
CDR-H1
Residues 31-35
NYGVT
VH
of SEQ ID
NO.: 49
S4-6
CDR-H2
Residues 50-65
MIWADGSTHYASSVKG
VH
of SEQ ID
NO.: 49
S4-6
CDR-H3
Residues 98-106
EWQHGPVAY
VH
of SEQ ID
NO.: 49
S4-18
SEQ ID NO.: 50
EVQLVESGGGLVQPGGSLRLSCAASGFTFS
VH
NYGVTWVRQAPGKGLEWVSMIWADGSTHYA
DSVKSRFTISRDNSKNTLYLQMNSLRAEDT
AVYYCAREWQHGPLAYWGQGTLVTVSS
S4-18
CDR-H1
Residues 31-35
NYGVT
VH
of SEQ ID
NO.: 50
S4-18
CDR-H2
Residues 50-65
MIWADGSTHYADSVKS
VH
of SEQ ID
NO.: 50
S4-18
CDR-H3
Residues 98-106
EWQHGPLAY
VH
of SEQ ID
NO.: 50
S4-31
SEQ ID NO.:51
EVQLVESGGGLVQPGGSLRLSCAASGFTFS
VH
NYGVQWVRQAPGKGLEWVSGIGADGSTAYA
SSLKGRFTISRDNSKNTLYLQMNSLRAEDT
AVYYCAREWQHSGLAYWGQGTLVTVSS
S4-31
CDR-H1
Residues 31-35
NYGVQ
VH
of SEQ ID
NO.: 51
S4-31
CDR-H2
Residues 50-65
GIGADGSTAYASSLKG
VH
of SEQ ID
NO.: 51
S4-31
CDR-H3
Residues 98-106
EWQHSGLAY
VH
of SEQ ID
NO.: 51
S4-34
SEQ ID NO.: 52
EVQLVESGGGLVQPGGSLRLSCAASGFTFS
VH
NYGVSWVRQAPGKGLEWVSMIWADGSTHYA
DTVKGRFTISRDNSKNTLYLQMNSLRAEDT
AVYYCAREWQHGPLAYWGQGTLVTVSS
S4-34
CDR-H1
Residues 31-35
NYGVS
VH
of SEQ ID
NO.: 52
S4-34
CDR-H2
Residues 50-65
MIWADGSTHYADTVKG
VH
of SEQ ID
NO.: 52
S4-34
CDR-H3
Residues 98-106
EWQHGPLAY
VH
of SEQ ID
NO.: 52
S4-74
SEQ ID NO.: 53
EVQLVESGGGLVQPGGSLRLSCAASGFTFS
VH
NYGVTWVRQAPGKGLEWVSMIWADGSTHYA
DTVKGRFTISRDNSKNTLYLQMNSLRAEDT
AVYYCAREWQHGPLAYWGQGTLVTVSS
S4-74
CDR-H1
Residues 31-35
NYGVT
VH
of SEQ ID
NO.: 53
S4-74
CDR-H2
Residues 50-65
MIWADGSTHYADTVKG
VH
of SEQ ID
NO.: 53
S4-74
CDR-H3
Residues 98-106
EWQHGPLAY
VH
of SEQ ID
NO.: 53
S4-12
SEQ ID NO.: 54
EVQLVESGGGLVQPGGSLRLSCAASGFTFS
VH
NYGVTWVRQAPGKGLEWVSMIWADGSTHYA
SSVKGRFTISRDNSKNTLYLQMNSLRAEDT
AVYYCAREWQHGPVAYWGQGTLVTVSS
S4-12
CDR-H1
Residues 31-35
NYGVT
VH
of SEQ ID
NO.: 54
S4-12
CDR-H2
Residues 50-65
MIWADGSTHYASSVKG
VH
of SEQ ID
NO.: 54
S4-12
CDR-H3
Residues 98-106
EWQHGPVAY
VH
of SEQ ID
NO.: 54
S4-54
SEQ ID NO.: 55
EVQLVESGGGLVQPGGSLRLSCAASGFTFS
VH
NYGVTWVRQAPGKGLEWVSMIWADGSTHYA
SSVKGRFTISRDNSKNTLYLQMNSLRAEDT
AVYYCAREWQHGPVAYWGQGTLVTVSS
S4-54
CDR-H1
Residues 31-35
NYGVT
VH
of SEQ ID
NO.: 55
S4-54
CDR-H2
Residues 50-65
MIWADGSTHYASSVKG
VH
of SEQ ID
NO.: 55
S4-54
CDR-H3
Residues 98-106
EWQHGPVAY
VH
of SEQ ID
NO.: 55
S4-17
SEQ ID NO.: 56
EVQLVESGGGLVQPGGSLRLSCAASGFTFS
VH
NYGVTWVRQAPGKGLEWVSMIWADGSTHYA
SSVKGRFTISRDNSKNTLYLQMNSLRAEDT
AVYYCAREWQHGPVAYWGQGTLVTVSS
S4-17
CDR-H1
Residues 31-35
NYGVT
VH
of SEQ ID
NO.: 56
S4-17
CDR-H2
Residues 50-65
MIWADGSTHYASSVKG
VH
of SEQ ID
NO.: 56
S4-17
CDR-H3
Residues 98-106
EWQHGPVAY
VH
of SEQ ID
NO.: 56
S4-40
SEQ ID NO.: 57
EVQLVESGGGLVQPGGSLRLSCAASGFTFS
VH
NYGVTWVRQAPGKGLEWVSMIWADGSTHYA
SSVKGRFTISRDNSKNTLYLQMNSLRAEDT
AVYYCAREWQHGPVAYWGQGTLVTVSS
S4-40
CDR-H1
Residues 31-35
NYGVT
VH
of SEQ ID
NO.: 57
S4-40
CDR-H2
Residues 50-65
MIWADGSTHYASSVKG
VH
of SEQ ID
NO.: 57
S4-40
CDR-H3
Residues 98-106
EWQHGPVAY
VH
of SEQ ID
NO.: 57
S4-24
SEQ ID NO.: 58
EVQLVESGGGLVQPGGSLRLSCAASGFTFS
VH
NYGVTWVRQAPGKGLEWVSMIWADGSTHYA
SSVKGRFTISRDNSKNTLYLQMNSLRAEDT
AVYYCAREWQHGPVAYWGQGTLVTVSS
S4-24
CDR-H1
Residues 31-35
NYGVT
VH
of SEQ ID
NO.: 58
S4-24
CDR-H2
Residues 50-65
MIWADGSTHYASSVKG
VH
of SEQ ID
NO.: 58
S4-24
CDR-H3
Residues 98-106
EWQHGPVAY
VH
of SEQ ID
NO.: 58
Table 7 provides VL sequences of IgG converted clones for Humanized anti-TNF MAK-195 antibodies as discussed in detail in Example 2.
TABLE 7
Humanized anti-TNF MAK-195 Ab VL sequences of
IgG converted clones
Sequence
Protein region
123456789012345678901234567890
hMAK195
SEQ ID NO.: 59
DIQMTQSPSSLSASVGDRVTITCKASQAVS
VL.1
SAVAWYQQKPGKAPKLLIYWASTRHTGVPS
VL
RFSGSGSGTDFTLTISSLQPEDFATYYCQQ
HYSTPFTFGQGTKLEIKR
hMAK195
CDR-L1
Residues 24-34
KASQAVSSAVA
VL.1
of SEQ ID
VL
NO.: 59
hMAK195
CDR-L2
Residues 50-56
WASTRHT
VL.1
of SEQ ID
VL
NO.: 59
hMAK195
CDR-L3
Residues 89-97
QQHYSTPFT
VL.1
of SEQ ID
VL
NO.: 59
S4-24
SEQ ID NO.: 60
DIQMTQSPSSLSASVGDRVTITCRASQLVS
VL
SAVAWYQQKPGKAPKLLIYWASTLHTGVPS
RFSGSGSGTDFTLTISSLQPEDFATYYCQQ
HYRTPFTFGQGTKLEIKR
S4-24
CDR-L1
Residues 24-34
RASQLVSSAVA
VL
of SEQ ID
NO.: 60
S4-24
CDR-L2
Residues 50-56
WASTLHT
VL
of SEQ ID
NO.: 60
S4-24
CDR-L3
Residues 89-97
QQHYRTPFT
VL
of SEQ ID
NO.: 60
S4-40
SEQ ID NO.: 61
DIQMTQSPSSLSASVGDRVTITCRASQLVS
VL
SAVAWYQQKPGKAPKLLIYWASTRHSGVPS
RFSGSGSGTDFTLTISSLQPEDFATYYCQQ
HYRTPFSFGQGTKLEIKR
S4-40
CDR-L1
Residues 24-34
RASQLVSSAVA
VL
of SEQ ID
NO.: 61
S4-40
CDR-L2
Residues 50-56
WASTRHS
VL
of SEQ ID
NO.: 61
S4-40
CDR-L3
Residues 89-97
QQHYRTPFS
VL
of SEQ ID
NO.: 61
S4-17
SEQ ID NO.: 62
DIQMTQSPSSLSASVGDRVTITCRASQLVS
VL
SAVAWYQQKPGKAPKLLIYWASTRHSGVPS
RFSGSGSGTDFTLTISSLQPEDFATYYCQQ
HYRTPFTFGQGTKLEIKR
S4-17
CDR-L1
Residues 24-34
RASQLVSSAVA
VL
of SEQ ID
NO.: 62
S4-17
CDR-L2
Residues 50-56
WASTRHS
VL
of SEQ ID
NO.: 62
S4-17
CDR-L3
Residues 89-97
QQHYRTPFT
VL
of SEQ ID
NO.: 62
S4-54
SEQ ID NO.: 63
DIQMTQSPSSLSASVGDRVTITCRASQLVS
VL
SAVAWYQQKPGKAPKLLIYWASARHTGVPS
RFSGSGSGTDFTLTISSLQPEDFATYYCQQ
HYKTPFSFGQGTKLEIKR
S4-54
CDR-L1
Residues 24-34
RASQLVSSAVA
VL
of SEQ ID
NO.: 63
S4-54
CDR-L2
Residues 50-56
WASARHT
VL
of SEQ ID
NO.: 63
S4-54
CDR-L3
Residues 89-97
QQHYKTPFS
VL
of SEQ ID
NO.: 63
S4-12
SEQ ID NO.: 64
DIQMTQSPSSLSASVGDRVTITCRASQLVS
VL
SAVAWYQQKPGKAPKLLIYWASARHTGVPS
RFSGSGSGTDFTLTISSLQPEDFATYYCQQ
HYKTPFTFGQGTKLEIKR
S4-12
CDR-L1
Residues 24-34
RASQLVSSAVA
VL
of SEQ ID
NO.: 64
S4-12
CDR-L2
Residues 50-56
WASARHT
VL
of SEQ ID
NO.: 64
S4-12
CDR-L3
Residues 89-97
QQHYKTPFT
VL
of SEQ ID
NO.: 64
S4-74
SEQ ID NO.: 65
DIQMTQSPSSLSASVGDRVTITCRASQLVS
VL
SAVAWYQQKPGKAPKLLIYWASARHTGVPS
RFSGSGSGTDFTLTISSLQPEDFATYYCQQ
HYRTPFTFGQGTKLEIKR
S4-74
CDR-L1
Residues 24-34
RASQLVSSAVA
VL
of SEQ ID
NO.: 65
S4-74
CDR-L2
Residues 50-56
WASARHT
VL
of SEQ ID
NO.: 65
S4-74
CDR-L3
Residues 89-97
QQHYRTPFT
VL
of SEQ ID
NO.: 65
S4-34
SEQ ID NO.: 66
DIQMTQSPSSLSASVGDRVTITCRASQLVS
VL
SAVAWYQQKPGKAPKLLIYWASTRHTGVPS
RFSGSGSGTDFTLTISSLQPEDFATYYCQQ
HYRTPFTFGQGTKLEIKR
S4-34
CDR-L1
Residues 24-34
RASQLVSSAVA
VL
of SEQ ID
NO.: 66
S4-34
CDR-L2
Residues 50-56
WASTRHT
VL
of SEQ ID
NO.: 66
S4-34
CDR-L3
Residues 89-97
QQHYRTPFT
VL
of SEQ ID
NO.: 66
S4-31
SEQ ID NO.: 67
DIQMTQSPSSLSASVGDRVTITCRASQGVS
VL
SALAWYQQKPGKAPKLLIYWASALHSGVPS
RFSGSGSGTDFTLTISSLQPEDFATYYCQQ
HYSAPFTFGQGTKLEIKR
S4-31
CDR-L1
Residues 24-34
RASQGVSSALA
VL
of SEQ ID
NO.: 67
S4-31
CDR-L2
Residues 50-56
WASALHS
VL
of SEQ ID
NO.: 67
S4-31
CDR-L3
Residues 89-97
QQHYSAPFT
VL
of SEQ ID
NO.: 67
S4-18
SEQ ID NO.: 68
DIQMTQSPSSLSASVGDRVTITCRASQLVS
VL
SAVAWYQQKPGKAPKLLIYWASTLHSGVPS
RFSGSGSGTDFTLTISSLQPEDFATYYCQQ
HYSTPFTFGQGTKLEIKR
S4-18
CDR-L1
Residues 24-34
RASQLVSSAVA
VL
of SEQ ID
NO.: 68
S4-18
CDR-L2
Residues 50-56
WASTLHS
VL
of SEQ ID
NO.: 68
S4-18
CDR-L3
Residues 89-97
QQHYSTPFT
VL
of SEQ ID
NO.: 68
S4-6
SEQ ID NO.: 69
DIQMTQSPSSLSASVGDRVTITCKASQLVS
VL
SAVAWYQQKPGKAPKLLIYWASTRHTGVPS
RFSGSGSGTDFTLTISSLQPEDFATYYCQQ
HYSTPFTFGQGTKLEIKR
S4-6
CDR-L1
Residues 24-34
KASQLVSSAVA
VL
of SEQ ID
NO.: 69
S4-6
CDR-L2
Residues 50-56
WASTRHT
VL
of SEQ ID
NO.: 69
S4-6
CDR-L3
Residues 89-97
QQHYSTPFT
VL
of SEQ ID
NO.: 69
S4-55
SEQ ID NO.: 70
DIQMTQSPSSLSASVGDRVTITCKASQLVS
VL
SAVAWYQQKPGKAPKLLIYWASTLHTGVPS
RFSGSGSGTDFTLTISSLQPEDFATYYCQQ
HYRTPFTFGQGTKLEIKR
S4-55
CDR-L1
Residues 24-34
KASQLVSSAVA
VL
of SEQ ID
NO.: 70
S4-55
CDR-L2
Residues 50-56
WASTLHT
VL
of SEQ ID
NO.: 70
S4-55
CDR-L3
Residues 89-97
QQHYRTPFT
VL
of SEQ ID
NO.: 70
S4-63
SEQ ID NO.: 71
DIQMTQSPSSLSASVGDRVTITCKASQKVS
VL
SALAWYQQKPGKAPKLLIYWASALHSGVPS
RFSGSGSGTDFTLTISSLQPEDFATYYCQQ
HYRPPFTFGQGTKLEIKR
S4-63
CDR-L1
Residues 24-34
KASQKVSSALA
VL
of SEQ ID
NO.: 71
S4-63
CDR-L2
Residues 50-56
WASALHS
VL
of SEQ ID
NO.: 71
S4-63
CDR-L3
Residues 89-97
QQHYRPPFT
VL
of SEQ ID
NO.: 71
S4-50
SEQ ID NO.: 72
DIQMTQSPSSLSASVGDRVTITCKASQLVS
VL
SAVAWYQQKPGKAPKLLIYWASALHTGVPS
RFSGSGSGTDFTLTISSLQPEDFATYYCQQ
HYSSPYTFGQGTKLEIKR
S4-50
CDR-L1
Residues 24-34
KASQLVSSAVA
VL
of SEQ ID
NO.: 72
S4-50
CDR-L2
Residues 50-56
WASALHT
VL
of SEQ ID
NO.: 72
S4-50
CDR-L3
Residues 89-97
QQHYSSPYT
VL
of SEQ ID
NO.: 72
S4-19
SEQ ID NO.: 73
DIQMTQSPSSLSASVGDRVTITCKASQLVS
VL
SAVAWYQQKPGKAPKLLIYWASTLHTGVPS
RFSGSGSGTDFTLTISSLQPEDFATYYCQQ
HYRTPFTFGQGTKLEIKR
S4-19
CDR-L1
Residues 24-34
KASQLVSSAVA
VL
of SEQ ID
NO.: 73
S4-19
CDR-L2
Residues 50-56
WASTLHT
VL
of SEQ ID
NO.: 73
S4-19
CDR-L3
Residues 89-97
QQHYRTPFT
VL
of SEQ ID
NO.: 73
C. Individual hMAK-199 Sequences from Converted Clones
Table 8 provides VH sequences of humanized anti-TNF MAK-199 converted clones as discussed in detail in Example 3.
TABLE 8
Humanized Anti-TNF MAK-199 Ab VH sequences of
IgG converted clones
Sequence
Protein region
123456789012345678901234567890
J662M2S3
SEQ ID NO.: 74
EVQLVQSGAEVKKPGASVKVSCKASGYTFA
#10 VH
NYGIIWVRQAPGQGLEWMGWINTYTGKPTY
AQKFQGRVTMTTDTSTSTAYMELSSLRSED
TAVYYCARKLFTTMDVTDNAMDYWGQGTTV
TVSS
J662M2S3#
CDR-H1
Residues 31-35
NYGII
10 VH
of SEQ ID
NO.: 74
J662M2S3#
CDR-H2
Residues 50-66
WINTYTGKPTYAQKFQG
10 VH
of SEQ ID
NO.: 74
J662M2S3#
CDR-H3
Residues 99-112
RASQDISQYLN
10 VH
of SEQ ID
NO.: 74
J662M2S3#
SEQ ID NO.: 75
EVQLVQSGAEVKKPGASVKVSCKASGYTFN
13 VH
NYGIIWVRQAPGQGLEWMGWINTYTGKPTY
AQKLQGRVTMTTDTSTSTAYMELSSLRSED
TAVYFCARKLFNTVDVTDNAMDYWGQGTTV
TVSS
J662M2S3#
CDR-H1
Residues 31-35
NYGII
13 VH
of SEQ ID
NO.: 75
J662M2S3#
CDR-H2
Residues 50-66
WINTYTGKPTYAQKLQG
13 VH
of SEQ ID
NO.: 75
J662M2S3#
CDR-H3
Residues 99-112
KLFNTVDVTDNAMD
13 VH
of SEQ ID
NO.: 75
J662M2S3#
SEQ ID NO.: 76
EVQLVQSGAEVKKPGASVKVSCKASGYTFN
15 VH
NYGIIWVRQAPGQGLEWMGWINTYTGVPTY
AQKFQGRVTMTTDTSTSTAYMELSSLRSED
TAVYYCARKLFNTVDVTDNAMDYWGQGTTV
TVSS
J662M2S3#
CDR-H1
Residues 31-35
NYGII
15 VH
of SEQ ID
NO.: 76
J662M2S3#
CDR-H2
Residues 50-66
WINTYTGVPTYAQKFQG
15 VH
of SEQ ID
NO.: 76
J662M2S3#
CDR-H3
Residues 99-112
KLFNTVDVTDNAMD
15 VH
of SEQ ID
NO.: 76
J662M2S3#
SEQ ID NO.: 77
EVQLVQSGAEVKKPGASVKVSCKASGYTFN
16 VH
NYGIIWVRQAPGQGLEWMGWINTYTGKPTY
AQKFQGRVTMTTDTSTSTAYMELSSLRSED
TAVYYCARKLFNTVAVTDNAMDYWGQGTTV
TVSS
J662M2S3#
CDR-H1
Residues 31-35
NYGII
16 VH
of SEQ ID
NO.: 77
J662M2S3#
CDR-H2
Residues 50-66
WINTYTGKPTYAQKFQG
16 VH
of SEQ ID
NO.: 77
J662M2S3#
CDR-H3
Residues 99-112
KLFNTVAVTDNAMD
16 VH
of SEQ ID
NO.: 77
J662M2S3#
SEQ ID NO.: 78
EVQLVQSGAEVKKPGASVKVSCKASGYTFR
21 VH
NYGIIWVRQAPGQGLEWMGWINTYTGKPTY
AQKFQGRVTMTTDTSTSTAYMELSSLRSED
TAVYFCARKLFTTVDVTDNAMDYWGQGTTV
TVSS
J662M2S3#
CDR-H1
Residues 31-35
NYGII
21 VH
of SEQ ID
NO.: 78
J662M2S3#
CDR-H2
Residues 50-66
WINTYTGKPTYAQKFQG
21 VH
of SEQ ID
NO.: 78
J662M2S3#
CDR-H3
Residues 99-112
KLFTTVDVTDNAMD
21 VH
of SEQ ID
NO.: 78
J662M2S3#
SEQ ID NO.: 79
EVQLVQSGAEVKKPGASVKVSCKASGYTFN
34 VH
NYGINWVRQAPGQGLEWMGWINTYTGKPTY
AQKFQGRVTMTTDTSTSTAYMELSSLRSED
TAVYFCARKFRNTVAVTDYAMDYWGQGTTV
TVSS
J662M2S3#
CDR-H1
Residues 31-35
NYGIN
34 VH
of SEQ ID
NO.: 79
J662M2S3#
CDR-H2
Residues 50-66
WINTYTGKPTYAQKFQG
34 VH
of SEQ ID
NO.: 79
J662M2S3#
CDR-H3
Residues 99-112
KFRNTVAVTDYAMD
34 VH
of SEQ ID
NO.: 79
J662M2S3#
SEQ ID NO.: 80
EVQLVQSGAEVKKPGASVKVSCKASGYTFR
36 VH
NYGITWVRQAPGQGLEWMGWINTYTGKPTY
AQKFQGRVTMTTDTSTSTAYMELSSLRSED
TAVYFCARKLFTTMDVTDNAMDYWGQGTTV
TVSS
J662M2S3#
CDR-H1
Residues 31-35
NYGIT
36 VH
of SEQ ID
NO.: 80
J662M2S3#
CDR-H2
Residues 50-66
WINTYTGKPTYAQKFQG
36 VH
of SEQ ID
NO.: 80
J662M2S3#
CDR-H3
Residues 99-112
KLFTTMDVTDNAMD
36 VH
of SEQ ID
NO.: 80
J662M2S3#
SEQ ID NO.: 81
EVQLVQSGAEVKKPGASVKVSCKASGYTFA
45 VH
NYGIIWVRQAPGQGLEWMGWINTYTGKPTY
AQKFQGRVTMTTDTSTSTAYMELSSLRSED
TAVYYCARKLFTTMDVTDNAMDYWGQGTTV
TVSS
J662M2S3#
CDR-H1
Residues 31-35
NYGII
45 VH
of SEQ ID
NO.: 81
J662M2S3#
CDR-H2
Residues 50-66
WINTYTGKPTYAQKFQG
45 VH
of SEQ ID
NO.: 81
J662M2S3#
CDR-H3
Residues 99-112
KLFTTMDVTDNAMD
45 VH
of SEQ ID
NO.: 81
J662M2S3#
SEQ ID NO.: 82
EVQLVQSGAEVKKPGASVKVSCKASGYTFS
58 VH
NYGINWVRQAPGQGLEWMGWINTYTGQPSY
AQKFQGRVTMTTDTSTSTAYMELSSLRSED
TAVYYCARKLFKTEAVTDYAMDYWGQGTTV
TVSS
J662M2S3#
CDR-H1
Residues 31-35
NYGIN
58 VH
of SEQ ID
NO.: 82
J662M2S3#
CDR-H2
Residues 50-66
WINTYTGQPSYAQKFQG
58 VH
of SEQ ID
NO.: 82
J662M2S3#
CDR-H3
Residues 99-112
KLFKTEAVTDYAMD
58 VH
of SEQ ID
NO.: 82
J662M2S3#
SEQ ID NO.: 83
EVQLVQSGAEVKKPGASVKVSCKASGYTFN
72 VH
NYGIIWVRQAPGQGLEWMGWINTYSGKPTY
AQKFQGRVTMTTDTSTSTAYMELSSLRSED
TAVYFCARKLFTTMDVTDNAMDYWGQGTTV
TVSS
J662M2S3#
CDR-H1
Residues 31-35
NYGII
72 VH
of SEQ ID
NO.: 83
J662M2S3#
CDR-H2
Residues 50-66
WINTYSGKPTYAQKFQG
72 VH
of SEQ ID
NO.: 83
J662M2S3#
CDR-H3
Residues 99-112
KLFTTMDVTDNAMD
72 VH
of SEQ ID
NO.: 83
Table 9 provides VL sequences of humanized anti-TNF MAK-199 converted clones as discussed in detail in Example 3.
TABLE 9
Humanized Anti-TNF MAK-199 Ab VL sequences of
IgG converted clones
Sequence
Protein region
123456789012345678901234567890
J662M2S3#
SEQ ID
DIQMTQSPSSLSASVGDRVTITCRASQDIS
10 VL
NO.: 84
QYLNWYQQKPGKAPKLLIYYTSRLQSGVPS
RFSGSGSGTDFTLTISSLQPEDFATYFCQQ
GNTWPPTFGQGTKLEIK
J662M2S3#10
CDR-L1
Residues 24-34
RASQDISQYLN
VL
of SEQ ID
NO.: 84
J662M2S3#10
CDR-L2
Residues 50-56
YTSRLQS
VL
of SEQ ID
NO.: 84
J662M2S3#10
CDR-L3
Residues 89-97
QQGNTWPPT
VL
of SEQ
ID NO.: 84
J662M2S3#13
SEQ ID
DIQMTQSPSSLSASVGDRVTITCRASQDIS
VL
NO.: 85
NYLNWYQQKPGKAPKLLIYYTSRLQSGVPS
RFSGSGSGTDYTLTISSLQPEDFATYFCQQ
GNSWPPTFGQGTKLEIK
J662M2S3#13
CDR-L1
Residues 24-34
RASQDISNYLN
VL
of SEQ ID
NO.: 85
J662M2S3#13
CDR-L2
Residues 50-56
YTSRLQS
VL
of SEQ ID
NO.: 85
J662M2S3#13
CDR-L3
Residues 89-97
QQGNSWPPT
VL
of SEQ
ID NO.: 85
J662M2S3#15
SEQ ID
DIQMTQSPSSLSASVGDRVTITCRASQDIY
VL
NO.: 86
NYLNWYQQKPGKAPKLLIYYTSRLQSGVPS
RFSGSGSGTDYTLTISSLQPEDFATYFCQQ
GNTQPPTFGQGTKLEIK
J662M2S3#15
CDR-L1
Residues 24-34
RASQDIYNYLN
VL
of SEQ ID
NO.: 86
J662M2S3#15
CDR-L2
Residues 50-56
YTSRLQS
VL
of SEQ ID
NO.: 86
J662M2S3#15
CDR-L3
Residues 89-97
QQGNTQPPT
VL
of SEQ
ID NO.: 86
J662M2S3#16
SEQ ID
DIQMTQSPSSLSASVGDRVTITCRASQDIE
VL
NO.: 87
NYLNWYQQKPGKAPKLLIYYTSRLQSGVPS
RFSGSGSGTDFTLTISSLQPEDFATYFCQQ
GNTQPPTFGQGTKLEIK
J662M2S3#16
CDR-L1
Residues 24-34
RASQDIENYLN
VL
of SEQ ID
NO.: 87
J662M2S3#16
CDR-L2
Residues 50-56
YTSRLQS
VL
of SEQ ID
NO.: 87
J662M2S3#16
CDR-L3
Residues 89-97
QQGNTQPPT
VL
of SEQ
ID NO.: 87
J662M2S3#21
SEQ ID
DIQMTQSPSSLSASVGDRVTITCRASQDIS
VL
NO.: 88
NYLNWYQQKPGKAPKLLIYYTSRLQSGVPS
RFSGSGSGTDYTLTISSLQPEDFATYFCQQ
GNTWPPTFGQGTKLEIK
J662M2S3#21
CDR-L1
Residues 24-34
RASQDISNYLN
VL
of SEQ ID
NO.: 88
J662M2S3#21
CDR-L2
Residues 50-56
YTSRLQS
VL
of SEQ ID
NO.: 88
J662M2S3#21
CDR-L3
Residues 89-97
QQGNTWPPT
VL
of SEQ
ID NO.: 88
J662M2S3#34
SEQ ID
DIQMTQSPSSLSASVGDRVTITCRASQDIY
VL
NO.: 89
DVLNWYQQKPGKAPKLLIYYASRLQSGVPS
RFSGSGSGTDFTLTISSLQPEDFATYYCQQ
GITLPPTFGQGTKLEIK
J662M2S3#34
CDR-L1
Residues 24-34
RASQDIYDVLN
VL
of SEQ ID
NO.: 89
J662M2S3#34
CDR-L2
Residues 50-56
YASRLQS
VL
of SEQ ID
NO.: 89
J662M2S3#34
CDR-L3
Residues 89-97
QQGITLPPT
VL
of SEQ
ID NO.: 89
J662M2S3#36
SEQ ID
DIQMTQSPSSLSASVGDRVTITCRASQDIS
VL
NO.: 90
NYLNWYQQKPGKAPKLLIYYTSRLQSGVPS
RFSGSGSGTDYTLTISSLQPEDFATYFCQQ
GNTWPPTFGQGTKLEIK
J662M2S3#36
CDR-L1
Residues 24-34
RASQDISNYLN
VL
of SEQ ID
NO.: 90
J662M2S3#36
CDR-L2
Residues 50-56
YTSRLQS
VL
of SEQ ID
NO.: 90
J662M2S3#36
CDR-L3
Residues 89-97
QQGNTWPPT
VL
of SEQ
ID NO.: 90
J662M2S3#45
SEQ ID
DIQMTQSPSSLSASVGDRVTITCRASQDIS
VL
NO.: 91
QYLNWYQQKPGKAPKLLIYYTSRLQSGVPS
RFSGSGSGTDFTLTISSLQPEDFATYFCQQ
GNTWPPTFGQGTKLEIK
J662M2S3#45
CDR-L1
Residues 24-34
RASQDISQYLN
VL
of SEQ ID
NO.: 91
J662M2S3#45
CDR-L2
Residues 50-56
YTSRLQS
VL
of SEQ ID
NO.: 91
J662M2S3#45
CDR-L3
Residues 89-97
QQGNTWPPT
VL
of SEQ
ID NO.: 91
J662M2S3#58
SEQ ID
DIQMTQSPSSLSASVGDRVTITCRASQNIY
VL
NO.: 92
NVLNWYQQKPGKAPKLLIYYASRLQSGVPS
RFSGSGSGTDFTLTISSLQPEDFATYFCQQ
GNTMPPTFGQGTKLEIK
J662M2S3#58
CDR-L1
Residues 24-34
RASQNIYNVLN
VL
of SEQ ID
NO.: 92
J662M2S3#58
CDR-L2
Residues 50-56
YASRLQS
VL
of SEQ ID
NO.: 92
J662M2S3#58
CDR-L3
Residues 89-97
QQGNTMPPT
VL
of SEQ
ID NO.: 92
J662M2S3#72
SEQ ID
DIQMTQSPSSLSASVGDRVTITCRASQDIS
VL
NO.: 93
NFLNWYQQKPGKAPKLLIYYTSRLQSGVPS
RFSGSGSGTDYTLTISSLQPEDFATYFCQQ
GNTQPPTFGQGTKLEIK
J662M2S3#72
CDR-L1
Residues 24-34
RASQDISNFLN
VL
of SEQ ID
NO.: 93
J662M2S3#72
CDR-L2
Residues 50-56
YTSRLQS
VL
of SEQ ID
NO.: 93
J662M2S3#72
CDR-L3
Residues 89-97
QQGNTQPPT
VL
of SEQ
ID NO.: 93
In an embodiment, the antigen binding domain comprises the VH region chosen from any one of SEQ ID NOs: 22, 24, 26, 28, 30, 32, 34-58, 74-83, 94-266, 478-486, 496-675, 738-762, 778-956, 1053-1062, 1073, 1075, and 1077, or one, two, or three CDRs therefrom. In another embodiment, the antigen binding domain comprises the VL region chosen from any one of SEQ ID NOs: 23, 25, 27, 29, 31, 33, 59-73, 84-93, 267-477, 487-495, 676-737, 763-777, 957-1052, 1063-1072, 1074, 1076, and 1078, or one, two, or three CDRs therefrom. In a particular embodiment, the antigen binding domain comprises a VH region and a VL region, for example, wherein the VH region comprises SEQ ID NOs: 22, 24, 26, 28, 30, 32, 34-58, 74-83, 94-266, 478-486, 496-675, 738-762, 778-956, 1053-1062, 1073, 1075, and 1077, or one, two, or three CDRs therefrom, and the VL region comprises SEQ ID NOs: 23, 25, 27, 29, 31, 33, 59-73, 84-93, 267-477, 487-495, 676-737, 763-777, 957-1052, 1063-1072, 1074, 1076, and 1078, or one, two, or three CDRs therefrom.
In an embodiment where the VH and/or the VL CDR sequences are provided above, the human acceptor framework comprises at least one amino acid sequence selected from: SEQ ID NOs: 6-21. In a particular embodiment, the human acceptor framework comprises an amino acid sequence selected from: SEQ IN NOs: 9, 10, 11, 12, 15, 16, 17, and 21. In another embodiment, the human acceptor framework comprises at least one framework region amino acid substitution, wherein the amino acid sequence of the framework is at least 65% identical to the sequence of the human acceptor framework and comprises at least 70 amino acid residues identical to the human acceptor framework. In another embodiment, the human acceptor framework comprises at least one framework region amino acid substitution at a key residue. The key residue selected from: a residue adjacent to a CDR; a glycosylation site residue; a rare residue; a residue capable of interacting with human TNF-α; a residue capable of interacting with a CDR; a canonical residue; a contact residue between heavy chain variable region and light chain variable region; a residue within a Vernier zone; and a residue in a region that overlaps between a Chothia-defined variable heavy chain CDR1 and a Kabat-defined first heavy chain framework. In an embodiment, the key residue is selected from: H1, H12, H24, H27, H29, H37, H48, H49, H67, H71, H73, H76, H78, L13, L43, L58, L70, and L80. In an embodiment, the VH mutation is selected from: Q1E, I12V, A24V, G27F, I29L, V29F F29L I37V, I48L, V48L, S49G, V67L, F67L, V71K, R71K, T73N, N76S, L78I, and F78I. In another embodiment, the VL mutation is selected from: V13L, A43S, I58V, E70D, and S80P. In an embodiment, the binding protein comprises two variable domains, wherein the two variable domains have amino acid sequences selected from: SEQ ID NOS: 22 and 23; 23 and 24; 24 and 25; 26 and 27; 28 and 29; 30 and 31; or 32 and 33.
III. Production of Binding Proteins and Binding Protein-Producing Cell Lines
In an embodiment, TNF-α binding proteins disclosed herein exhibit a high capacity to reduce or to neutralize TNF-α activity, e.g., as assessed by any one of several in vitro and in vivo assays known in the art. Alternatively, TNF-α binding proteins disclosed herein, also exhibit a high capacity to increase or agonize TNF-α activity.
In particular embodiments, the isolated binding protein, or antigen-binding portion thereof, binds human TNF-α, wherein the binding protein, or antigen-binding portion thereof, dissociates from human TNF-α with a koff rate constant of about 0.1 s−1 or less, as determined by surface plasmon resonance, such as 1×10−2 s−1 or less, 1×10−3 s−1 or less, 1×10−4 s−1 or less, 1×10−5 s−1 or less and 1×10−6 s−1 or less; or which inhibits human TNF-α activity with an IC50 of about 1×10−6 M or less, such as 1×10−7 M or less, 1×10−8 M or less, 1×10−9 M or less, 1×10−10 M or less and 1×10−11 M or less. In certain embodiments, the binding protein comprises a heavy chain constant region, such as an IgG1, IgG2, IgG3, IgG4, IgA, IgE, IgM or IgD constant region. In an embodiment, the heavy chain constant region is an IgG1 heavy chain constant region or an IgG4 heavy chain constant region. Furthermore, the binding protein can comprise a light chain constant region, either a kappa light chain constant region or a lambda light chain constant region. In another embodiment, the binding protein comprises a kappa light chain constant region. Alternatively, the binding protein portion can be, for example, a Fab fragment or a single chain Fv fragment.
Replacements of amino acid residues in the Fc portion to alter binding protein effector function are known in the art (See U.S. Pat. Nos. 5,648,260 and 5,624,821). The Fc portion of a binding protein mediates several important effector functions, e.g., cytokine induction, ADCC, phagocytosis, complement dependent cytotoxicity (CDC) and half-life/clearance rate of antibody and antigen-antibody complexes. In some cases these effector functions are desirable for therapeutic antibody but in other cases might be unnecessary or even deleterious, depending on the therapeutic objectives. Certain human IgG isotypes, particularly IgG1 and IgG3, mediate ADCC and CDC via binding to FcγRs and complement C1q, respectively. Neonatal Fc receptors (FcRn) are the critical components determining the circulating half-life of antibodies. In still another embodiment at least one amino acid residue is replaced in the constant region of the binding protein, for example the Fc region of the binding protein, such that effector functions of the binding protein are altered.
One embodiment provides a labeled binding protein wherein an antibody or antibody portion disclosed herein is derivatized or linked to another functional molecule (e.g., another peptide or protein). For example, a labeled binding protein disclosed herein can be derived by functionally linking an antibody or antibody portion disclosed herein (by chemical coupling, genetic fusion, noncovalent association or otherwise) to one or more other molecular entities, such as another antibody (e.g., a bispecific antibody or a diabody), a detectable agent, a cytotoxic agent, a pharmaceutical agent, and/or a protein or peptide that can mediate associate of the antibody or antibody portion with another molecule (such as a streptavidin core region or a polyhistidine tag).
Useful detectable agents with which an antibody or antibody portion disclosed herein may be derivatized include fluorescent compounds. Exemplary fluorescent detectable agents include fluorescein, fluorescein isothiocyanate, rhodamine, 5-dimethylamine-1-napthalenesulfonyl chloride, phycoerythrin and the like. An antibody may also be derivatized with detectable enzymes, such as alkaline phosphatase, horseradish peroxidase, glucose oxidase and the like. When an antibody is derivatized with a detectable enzyme, it is detected by adding additional reagents that the enzyme uses to produce a detectable reaction product. For example, when the detectable agent horseradish peroxidase is present, the addition of hydrogen peroxide and diaminobenzidine leads to a colored reaction product, which is detectable. An antibody may also be derivatized with biotin, and detected through indirect measurement of avidin or streptavidin binding.
Another embodiment of the disclosure provides a crystallized binding protein. In an embodiment, provided are crystals of whole TNF-α binding proteins and fragments thereof as disclosed herein, and formulations and compositions comprising such crystals. In one embodiment the crystallized binding protein has a greater half-life in vivo than the soluble counterpart of the binding protein. In another embodiment the binding protein retains biological activity after crystallization.
Crystallized binding protein disclosed herein may be produced according methods known in the art and as disclosed in PCT Publication WO 02/72636.
Another embodiment of the disclosure provides a glycosylated binding protein wherein the binding protein or antigen-binding portion thereof comprises one or more carbohydrate residues. Nascent in vivo protein production may undergo further processing, known as post-translational modification. In particular, sugar (glycosyl) residues may be added enzymatically, a process known as glycosylation. The resulting proteins bearing covalently linked oligosaccharide side chains are known as glycosylated proteins or glycoproteins. Protein glycosylation depends on the amino acid sequence of the protein of interest, as well as the host cell in which the protein is expressed. Different organisms may produce different glycosylation enzymes (e.g., glycosyltransferases and glycosidases), and have different substrates (nucleotide sugars) available. Due to such factors, protein glycosylation pattern, and composition of glycosyl residues, may differ depending on the host system in which the particular protein is expressed. Glycosyl residues useful in the disclosure may include, but are not limited to, glucose, galactose, mannose, fucose, n-acetylglucosamine and sialic acid. In an embodiment, the glycosylated binding protein comprises glycosyl residues such that the glycosylation pattern is human.
It is known to those skilled in the art that differing protein glycosylation may result in differing protein characteristics. For instance, the efficacy of a therapeutic protein produced in a microorganism host, such as yeast, and glycosylated utilizing the yeast endogenous pathway may be reduced compared to that of the same protein expressed in a mammalian cell, such as a CHO cell line. Such glycoproteins may also be immunogenic in humans and show reduced half-life in vivo after administration. Specific receptors in humans and other animals may recognize specific glycosyl residues and promote the rapid clearance of the protein from the bloodstream. Other adverse effects may include changes in protein folding, solubility, susceptibility to proteases, trafficking, transport, compartmentalization, secretion, recognition by other proteins or factors, antigenicity, or allergenicity. Accordingly, a practitioner may prefer a therapeutic protein with a specific composition and pattern of glycosylation, for example glycosylation composition and pattern identical, or at least similar, to that produced in human cells or in the species-specific cells of the intended subject animal.
Expressing glycosylated proteins different from that of a host cell may be achieved by genetically modifying the host cell to express heterologous glycosylation enzymes. Using techniques known in the art a practitioner may generate antibodies or antigen-binding portions thereof exhibiting human protein glycosylation. For example, yeast strains have been genetically modified to express non-naturally occurring glycosylation enzymes such that glycosylated proteins (glycoproteins) produced in these yeast strains exhibit protein glycosylation identical to that of animal cells, especially human cells (U.S. Pat. Nos. 7,449,308 and 7,029,872).
Further, it will be appreciated by one skilled in the art that a protein of interest may be expressed using a library of host cells genetically engineered to express various glycosylation enzymes, such that member host cells of the library produce the protein of interest with variant glycosylation patterns. A practitioner may then select and isolate the protein of interest with particular novel glycosylation patterns. In an embodiment, the protein having a particularly selected novel glycosylation pattern exhibits improved or altered biological properties.
IV. Uses of TNF-α Binding Proteins
Given their ability to bind to human TNF-α, e.g., the human TNF-α binding proteins, or portions thereof, disclosed herein can be used to detect TNF-α (e.g., in a biological sample, such as serum or plasma), using a conventional immunoassay, such as an enzyme linked immunosorbent assays (ELISA), an radioimmunoassay (RIA) or tissue immunohistochemistry. A method for detecting TNF-α in a biological sample is provided comprising contacting a biological sample with a binding protein, or binding protein portion, disclosed herein and detecting either the binding protein (or binding protein portion) bound to TNF-α or unbound binding protein (or binding protein portion), to thereby detect TNF-α in the biological sample. The binding protein is directly or indirectly labeled with a detectable substance to facilitate detection of the bound or unbound antibody. Suitable detectable substances include various enzymes, prosthetic groups, fluorescent materials, luminescent materials and radioactive materials. Examples of suitable enzymes include horseradish peroxidase, alkaline phosphatase, β-galactosidase, or acetylcholinesterase; examples of suitable prosthetic group complexes include streptavidin/biotin and avidin/biotin; examples of suitable fluorescent materials include umbelliferone, fluorescein, fluorescein isothiocyanate, rhodamine, dichlorotriazinylamine fluorescein, dansyl chloride or phycoerythrin; an example of a luminescent material includes luminol; and examples of suitable radioactive material include 3H, 14C, 35S, 90Y, 99Tc, 111In, 125I, 131I, 177Lu, 166Ho, or 153Sm.
Alternative to labeling the binding protein, human TNF-α can be assayed in biological fluids by a competition immunoassay utilizing rhTNF-α standards labeled with a detectable substance and an unlabeled human TNF-α binding protein. In this assay, the biological sample, the labeled rhTNF-α standards and the human TNF-α binding protein are combined and the amount of labeled rhTNF-α standard bound to the unlabeled binding protein is determined. The amount of human TNF-α in the biological sample is inversely proportional to the amount of labeled rhTNF-α standard bound to the TNF-α binding protein. Similarly, human TNF-α can also be assayed in biological fluids by a competition immunoassay utilizing rhTNF-α standards labeled with a detectable substance and an unlabeled human TNF-α binding protein.
In an embodiment, the binding proteins and binding protein portions disclosed herein are capable of neutralizing TNF-α activity, e.g., human TNF-α activity, both in vitro and in vivo. In another embodiment, the binding proteins and binding protein portions disclosed herein are capable of increasing or agonizing human TNF-α activity, e.g., human TNF-α activity. Accordingly, such binding proteins and binding protein portions disclosed herein can be used to inhibit or increase hTNF-α activity, e.g., in a cell culture containing hTNF-α, in human subjects or in other mammalian subjects having TNF-α with which a binding protein disclosed herein cross-reacts. In one embodiment, a method for inhibiting or increasing hTNF-α activity is provided comprising contacting hTNF-α with a binding protein or binding protein portion disclosed herein such that hTNF-α activity is inhibited or increased. For example, in a cell culture containing, or suspected of containing hTNF-α, a binding protein or binding protein portion disclosed herein can be added to the culture medium to inhibit or increase hTNF-α activity in the culture.
In another embodiment, a method is provided for reducing or increasing hTNF-α activity in a subject, advantageously from a subject suffering from a disease or disorder in which TNF-α-activity is detrimental or, alternatively, beneficial. Methods for reducing or increasing TNF-α activity in a subject suffering from such a disease or disorder is provided, which method comprises administering to the subject a binding protein or binding protein portion disclosed herein such that TNF-α activity in the subject is reduced or increased. In a particular embodiment, the TNF-α is human TNF-α, and the subject is a human subject. Alternatively, the subject can be a mammal expressing a TNF-α to which a binding protein provided is capable of binding. Still further the subject can be a mammal into which TNF-α has been introduced (e.g., by administration of TNF-α or by expression of a TNF-α transgene). A binding protein disclosed herein can be administered to a human subject for therapeutic purposes. Moreover, a binding protein disclosed herein can be administered to a non-human mammal expressing a TNF-α with which the binding protein is capable of binding for veterinary purposes or as an animal model of human disease. Regarding the latter, such animal models may be useful for evaluating the therapeutic efficacy of binding proteins disclosed herein (e.g., testing of dosages and time courses of administration).
The term “a disorder in which TNF-α activity is detrimental” includes diseases and other disorders in which the presence of TNF-α activity in a subject suffering from the disorder has been shown to be or is suspected of being either responsible for the pathophysiology of the disorder or a factor that contributes to a worsening of the disorder. Accordingly, a disorder in which TNF-α activity is detrimental is a disorder in which reduction of TNF-α activity is expected to alleviate the symptoms and/or progression of the disorder. Such disorders may be evidenced, for example, by an increase in the concentration of TNF-α in a biological fluid of a subject suffering from the disorder (e.g., an increase in the concentration of TNF-α in serum, plasma, synovial fluid, etc. of the subject), which can be detected, for example, using an anti-TNF-α antibody as described above. Non-limiting examples of disorders that can be treated with the binding proteins disclosed herein include those disorders discussed in the section below pertaining to pharmaceutical compositions of the antibodies disclosed herein.
Alternatively, the term “a disorder in which TNF-α activity is beneficial” include diseases and other disorders in which the presence of TNF-α activity in a subject suffering from the disorder has been shown to be or is suspected of being either beneficial for treating the pathophysiology of the disorder or a factor that contributes to a treatment of the disorder. Accordingly, a disorder in which TNF-α activity is beneficial is a disorder in which an increase of TNF-α activity is expected to alleviate the symptoms and/or progression of the disorder. Non-limiting examples of disorders that can be treated with the antibodies disclosed herein include those disorders discussed in the section below pertaining to pharmaceutical compositions of the antibodies disclosed herein.
V. Pharmaceutical Compositions
Pharmaceutical compositions are also provided comprising a binding protein, or antigen-binding portion thereof, disclosed herein and a pharmaceutically acceptable carrier. The pharmaceutical compositions comprising binding protein disclosed herein are for use in, but not limited to, diagnosing, detecting, or monitoring a disorder, in preventing, treating, managing, or ameliorating of a disorder or one or more symptoms thereof, and/or in research. In a specific embodiment, a composition comprises one or more binding proteins disclosed herein. In another embodiment, the pharmaceutical composition comprises one or more binding proteins disclosed herein and one or more prophylactic or therapeutic agents other than binding proteins disclosed herein for treating a disorder in which TNF-α activity is detrimental. In a particular embodiment, the prophylactic or therapeutic agents known to be useful for or having been or currently being used in the prevention, treatment, management, or amelioration of a disorder or one or more symptoms thereof. In accordance with these embodiments, the composition may further comprise of a carrier, diluent or excipient.
The binding proteins and binding protein-portions disclosed herein can be incorporated into pharmaceutical compositions suitable for administration to a subject. Typically, the pharmaceutical composition comprises a binding protein or binding protein portion disclosed herein and a pharmaceutically acceptable carrier. The term “pharmaceutically acceptable carrier” includes any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, and the like that are physiologically compatible. Examples of pharmaceutically acceptable carriers include one or more of water, saline, phosphate buffered saline, dextrose, glycerol, ethanol and the like, as well as combinations thereof. In many cases, isotonic agents, for example, sugars, polyalcohols such as mannitol, sorbitol, or sodium chloride in the composition, may be included. Pharmaceutically acceptable carriers may further comprise minor amounts of auxiliary substances such as wetting or emulsifying agents, preservatives or buffers, which enhance the shelf life or effectiveness of the binding protein or binding protein portion.
Various delivery systems are known and can be used to administer one or more binding proteins disclosed herein or the combination of one or more binding proteins disclosed herein and a prophylactic agent or therapeutic agent useful for preventing, managing, treating, or ameliorating a disorder or one or more symptoms thereof, e.g., encapsulation in liposomes, microparticles, microcapsules, recombinant cells capable of expressing the binding protein or binding protein fragment, receptor-mediated endocytosis (see, e. g., Wu and Wu (1987) J. Biol. Chem. 262:4429-4432), construction of a nucleic acid as part of a retroviral or other vector, etc. Methods of administering a prophylactic or therapeutic agent disclosed herein include, but are not limited to, parenteral administration (e.g., intradermal, intramuscular, intraperitoneal, intravenous and subcutaneous), epidural administration, intratumoral administration, and mucosal administration (e.g., intranasal and oral routes). In addition, pulmonary administration can be employed, e.g., by use of an inhaler or nebulizer, and formulation with an aerosolizing agent. In one embodiment, a binding protein disclosed herein, combination therapy, or a composition disclosed herein is administered using Alkermes AIR® pulmonary drug delivery technology (Alkermes, Inc., Cambridge, Mass.). In a specific embodiment, prophylactic or therapeutic agents disclosed herein are administered intramuscularly, intravenously, intratumorally, orally, intranasally, pulmonary, or subcutaneously. The prophylactic or therapeutic agents may be administered by any convenient route, for example by infusion or bolus injection, by absorption through epithelial or mucocutaneous linings (e.g., oral mucosa, rectal and intestinal mucosa, etc.) and may be administered together with other biologically active agents. Administration can be systemic or local.
In a specific embodiment, it may be desirable to administer the prophylactic or therapeutic agents disclosed herein locally to the area in need of treatment; this may be achieved by, for example, and not by way of limitation, local infusion, by injection, or by means of an implant, said implant being of a porous or non-porous material, including membranes and matrices, such as sialastic membranes, polymers, fibrous matrices (e.g., Tissuel®), or collagen matrices. In one embodiment, an effective amount of one or more binding proteins disclosed herein antagonists is administered locally to the affected area to a subject to prevent, treat, manage, and/or ameliorate a disorder or a symptom thereof. In another embodiment, an effective amount of one or more binding proteins disclosed herein is administered locally to the affected area in combination with an effective amount of one or more therapies (e.g., one or more prophylactic or therapeutic agents) other than an antibody disclosed herein of a subject to prevent, treat, manage, and/or ameliorate a disorder or one or more symptoms thereof.
In a specific embodiment, where the composition disclosed herein is a nucleic acid encoding a prophylactic or therapeutic agent, the nucleic acid can be administered in vivo to promote expression of its encoded prophylactic or therapeutic agent, by constructing it as part of an appropriate nucleic acid expression vector and administering it so that it becomes intracellular, e.g., by use of a retroviral vector (see U.S. Pat. No. 4,980,286), or by direct injection, or by use of microparticle bombardment (e.g., a gene gun; Biolistic, DuPont), or coating with lipids or cell-surface receptors or transfecting agents, or by administering it in linkage to a homeobox-like peptide which is known to enter the nucleus (see, e.g., Joliot et al. (1991) Proc. Natl. Acad. Sci. USA 88:1864-1868). Alternatively, a nucleic acid can be introduced intracellularly and incorporated within host cell DNA for expression by homologous recombination.
The method disclosed herein may comprise administration of a composition formulated for parenteral administration by injection (e.g., by bolus injection or continuous infusion). Formulations for injection may be presented in unit dosage form (e.g., in ampoules or in multi-dose containers) with an added preservative. The compositions may take such forms as suspensions, solutions or emulsions in oily or aqueous vehicles, and may contain formulatory agents such as suspending, stabilizing and/or dispersing agents. Alternatively, the active ingredient may be in powder form for constitution with a suitable vehicle (e.g., sterile pyrogen-free water) before use.
The methods disclosed herein may additionally comprise of administration of compositions formulated as depot preparations. Such long acting formulations may be administered by implantation (e.g., subcutaneously or intramuscularly) or by intramuscular injection. Thus, for example, the compositions may be formulated with suitable polymeric or hydrophobic materials (e.g., as an emulsion in an acceptable oil) or ion exchange resins, or as sparingly soluble derivatives (e.g., as a sparingly soluble salt).
The methods disclosed herein encompass administration of compositions formulated as neutral or salt forms. Pharmaceutically acceptable salts include those formed with anions such as those derived from hydrochloric, phosphoric, acetic, oxalic, tartaric acids, etc., and those formed with cations such as those derived from sodium, potassium, ammonium, calcium, ferric hydroxides, isopropylamine, triethylamine, 2-ethylamino ethanol, histidine, procaine, etc.
Generally, the ingredients of compositions are supplied either separately or mixed together in unit dosage form, for example, as a dry lyophilized powder or water free concentrate in a hermetically sealed container such as an ampoule or sachette indicating the quantity of active agent. Where the mode of administration is infusion, composition can be dispensed with an infusion bottle containing sterile pharmaceutical grade water or saline. Where the mode of administration is by injection, an ampoule of sterile water for injection or saline can be provided so that the ingredients may be mixed prior to administration.
In particular, it is also provided that one or more of the prophylactic or therapeutic agents, or pharmaceutical compositions disclosed herein is packaged in a hermetically sealed container such as an ampoule or sachette indicating the quantity of the agent. In one embodiment, one or more of the prophylactic or therapeutic agents, or pharmaceutical compositions disclosed herein is supplied as a dry sterilized lyophilized powder or water free concentrate in a hermetically sealed container and can be reconstituted (e.g., with water or saline) to the appropriate concentration for administration to a subject. In an embodiment, one or more of the prophylactic or therapeutic agents or pharmaceutical compositions disclosed herein is supplied as a dry sterile lyophilized powder in a hermetically sealed container at a unit dosage of at least 5 mg, at least 10 mg, at least 15 mg, at least 25 mg, at least 35 mg, at least 45 mg, at least 50 mg, at least 75 mg, or at least 100 mg. The lyophilized prophylactic or therapeutic agents or pharmaceutical compositions disclosed herein should be stored at between 2° C. and 8° C. in its original container and the prophylactic or therapeutic agents, or pharmaceutical compositions disclosed herein should be administered within 1 week, within 5 days, within 72 hours, within 48 hours, within 24 hours, within 12 hours, within 6 hours, within 5 hours, within 3 hours, or within 1 hour after being reconstituted. In an alternative embodiment, one or more of the prophylactic or therapeutic agents or pharmaceutical compositions disclosed herein is supplied in liquid form in a hermetically sealed container indicating the quantity and concentration of the agent. In an embodiment, the liquid form of the administered composition is supplied in a hermetically sealed container at least 0.25 mg/ml, at least 0.5 mg/ml, at least 1 mg/ml, at least 2.5 mg/ml, at least 5 mg/ml, at least 8 mg/ml, at least 10 mg/ml, at least 15 mg/kg, at least 25 mg/ml, at least 50 mg/ml, at least 75 mg/ml or at least 100 mg/ml. The liquid form should be stored at between 2° C. and 8° C. in its original container.
The binding proteins and binding protein-portions disclosed herein can be incorporated into a pharmaceutical composition suitable for parenteral administration. In an embodiment, the binding protein or binding protein-portions will be prepared as an injectable solution containing 0.1-250 mg/ml binding protein. The injectable solution can be composed of either a liquid or lyophilized dosage form in a flint or amber vial, ampoule or pre-filled syringe. The buffer can be L-histidine (1-50 mM), optimally 5-10 mM, at pH 5.0 to 7.0 (optimally pH 6.0). Other suitable buffers include but are not limited to, sodium succinate, sodium citrate, sodium phosphate or potassium phosphate. Sodium chloride can be used to modify the toxicity of the solution at a concentration of 0-300 mM (optimally 150 mM for a liquid dosage form). Cryoprotectants can be included for a lyophilized dosage form, principally 0-10% sucrose (optimally 0.5-1.0%). Other suitable cryoprotectants include trehalose and lactose. Bulking agents can be included for a lyophilized dosage form, principally 1-10% mannitol (optimally 2-4%). Stabilizers can be used in both liquid and lyophilized dosage forms, principally 1-50 mM L-Methionine (optimally 5-10 mM). Other suitable bulking agents include glycine, arginine, can be included as 0-0.05% polysorbate-80 (optimally 0.005-0.01%). Additional surfactants include but are not limited to polysorbate 20 and BRIJ surfactants.
Typical compositions are in the form of injectable or infusible solutions, such as compositions similar to those used for passive immunization of humans with other antibodies. Therapeutic compositions typically must be sterile and stable under the conditions of manufacture and storage. The composition can be formulated as a solution, microemulsion, dispersion, liposome, or other ordered structure suitable to high drug concentration. Sterile injectable solutions can be prepared by incorporating the active compound (i.e., binding protein or binding protein portion) in the required amount in an appropriate solvent with one or a combination of ingredients enumerated above, as required, followed by filtered sterilization. Generally, dispersions are prepared by incorporating the active compound into a sterile vehicle that contains a basic dispersion medium and the required other ingredients from those enumerated above. In the case of sterile, lyophilized powders for the preparation of sterile injectable solutions, the methods of preparation include vacuum drying and spray-drying that yields a powder of the active ingredient plus any additional desired ingredient from a previously sterile-filtered solution thereof. The proper fluidity of a solution can be maintained, for example, by the use of a coating such as lecithin, by the maintenance of the required particle size in the case of dispersion and by the use of surfactants. Prolonged absorption of injectable compositions can be brought about by including, in the composition, an agent that delays absorption, for example, monostearate salts and gelatin.
As will be appreciated by the skilled artisan, the route and/or mode of administration will vary depending upon the desired results. In certain embodiments, the active compound may be prepared with a carrier that will protect the compound against rapid release, such as a controlled release formulation, including implants, transdermal patches, and microencapsulated delivery systems. Biodegradable, biocompatible polymers can be used, such as ethylene vinyl acetate, polyanhydrides, polyglycolic acid, collagen, polyorthoesters, and polylactic acid. Many methods for the preparation of such formulations are patented or generally known to those skilled in the art. See, e.g., Sustained and Controlled Release Drug Delivery Systems, J. R. Robinson, ed., Marcel Dekker, Inc., New York, 1978.
Supplementary active compounds can also be incorporated into the compositions. In certain embodiments, a binding protein or binding protein portion disclosed herein is coformulated with and/or coadministered with one or more additional therapeutic agents that are useful for treating disorders in which TNF-α activity is detrimental. For example, an anti-hTNF-α antibody or antibody portion disclosed herein may be coformulated and/or coadministered with one or more additional antibodies that bind other targets (e.g., antibodies that bind other cytokines or that bind cell surface molecules). Furthermore, one or more binding proteins disclosed herein may be used in combination with two or more of the foregoing therapeutic agents. Such combination therapies may advantageously utilize lower dosages of the administered therapeutic agents, thus avoiding possible toxicities or complications associated with the various monotherapies.
In certain embodiments, a binding protein to TNF-α or fragment thereof is linked to a half-life extending vehicle known in the art. Such vehicles include, but are not limited to, the Fc domain, polyethylene glycol, and dextran. Such vehicles are described, e.g., in U.S. Pat. No. 6,660,843.
In a specific embodiment, nucleic acid sequences comprising nucleotide sequences encoding a binding protein disclosed herein or another prophylactic or therapeutic agent disclosed herein are administered to treat, prevent, manage, or ameliorate a disorder or one or more symptoms thereof by way of gene therapy. Gene therapy refers to therapy performed by the administration to a subject of an expressed or expressible nucleic acid. In this embodiment of the disclosure, the nucleic acids produce their encoded binding protein or prophylactic or therapeutic agent disclosed herein that mediates a prophylactic or therapeutic effect.
Any of the methods for gene therapy available in the art can be used according to the present disclosure.
TNF-α plays a critical role in the pathology associated with a variety of diseases involving immune and inflammatory elements, such as autoimmune diseases, particularly those assocated with inflammation, including Crohn's disease, psoriasis (including plaque psoriasis), arthritis (including rheumatoid arthritis, psoratic arthritis, osteoarthritis, or juvenile idiopathic arthritis), multiple sclerosis, systemic lupus erythematosus, and ankylosing spondylitis. Therefore, the binding proteins herein may be used to treat these disorders. In another embodiment, the disorder is a respiratory disorder; asthma; allergic and nonallergic asthma; asthma due to infection; asthma due to infection with respiratory syncytial virus (RSV); chronic obstructive pulmonary disease (COPD); a condition involving airway inflammation; eosinophilia; fibrosis and excess mucus production; cystic fibrosis; pulmonary fibrosis; an atopic disorder; atopic dermatitis; urticaria; eczema; allergic rhinitis; allergic enterogastritis; an inflammatory and/or autoimmune condition of the skin; an inflammatory and/or autoimmune condition of gastrointestinal organs; inflammatory bowel diseases (IBD); ulcerative colitis; an inflammatory and/or autoimmune condition of the liver; liver cirrhosis; liver fibrosis; liver fibrosis caused by hepatitis B and/or C virus; scleroderma; tumors or cancers; hepatocellular carcinoma; glioblastoma; lymphoma; Hodgkin's lymphoma; a viral infection; a bacterial infection; a parasitic infection; HTLV-1 infection; suppression of expression of protective type 1 immune responses, suppression of expression of a protective type 1 immune response during vaccination, neurodegenerative diseases, neuronal regeneration, and spinal cord injury.
It will be readily apparent to those skilled in the art that other suitable modifications and adaptations of the methods disclosed herein may be made using suitable equivalents without departing from the scope of the invention or the embodiments disclosed herein. Having now described the present disclosure in detail, the same will be more clearly understood by reference to the following examples, which are included for purposes of illustration only and are not intended to be limiting of the invention.
Fully human anti-human TNF monoclonal antibodies were isolated by in vitro display technologies from human antibody libraries by their ability to bind recombinant human TNF proteins. The amino acid sequences of the variable heavy (VH) and variable light (VL) chains were determined from DNA sequencing and listed in Table 10.
TABLE 10
Individual clones sequences
Protein
Sequence
region
SEQ ID NO:
123456789012345678901234567890
AE11-1 VH
22
EVQLVQSGAEVKKPGASVKVSCKASGYTFT
SYDVNWVRQATGQGLEWMGMNPNSGNTGY
AQKFQGRVTITADESTSTAYMELSSLRSED
TAVYYCAIFDSDYMDVWGKGTLVTVSS
AE11-1 VH
CDR-
Residues 31-35
SYDVN
H1
of SEQ ID
NO.: 22
AE11-1 VH
CDR-
Residues 50-66
WMNPNSGNTGYAQKFQG
H2
of SEQ ID
NO.: 22
AE11-1 VH
CDR-
Residues 99-106
FDSDYMDV
H3
of SEQ ID
NO.: 22
AE11-1 VL
23
SYELTQPPSVSLSPGQTARITCSGDALPKQ
YAYWYQQKPGQAPVLVIYKDTERPSGIPER
FSGSSSGTTVTLTISGAQAEDEADYYCQSA
DSSGTSWVFGGGTKLTVL
AE11-1 VL
CDR-
Residues 23-33
SGDALPKQYAY
L1
of SEQ ID
NO.: 23
AE11-1 VL
CDR-
Residues 49-55
KDTERPS
L2
of SEQ ID
NO.: 23
AE11-1 VL
CDR-
Residues 89-98
SADSSGTSWV
L3
of SEQ ID
NO.: 23
AE11-5 VH
24
EVQLVQSGAEVKKPGSSAKVSCKASGGTFS
SYAISWVRQAPGQGLEWMGGIIPILGTANY
AQKFLGRVTITADESTSTVYMELSSLRSED
TAVYYCARGLYYDPTRADYWGQGTLVTVSS
AE11-5 VH
CDR-
Residues 31-35
SYAIS
H1
of SEQ ID
NO.: 24
AE11-5 VH
CDR-
Residues 50-66
GIIPILGTANYAQKFLG
H2
of SEQ ID
NO.: 24
AE11-5 VH
CDR-
Residues 99-109
GLYYDPTRADY
H3
of SEQ ID
NO.: 24
AE11-5 VL
25
DIVMTQSPDFHSVTPKEKVTITCRASQSIG
SSLHWYQQKPDQSPKLLIRHASQSISGVPS
RFSGSGSGTDFTLTIHSLEAEDAATYYCHQ
SSSSPPPTFGQGTQVEIK
AE11-5 VL
CDR-
Residues 24-34
RASQSIGSSLH
L1
of SEQ ID
NO.: 25
AE11-5 VL
CDR-
Residues 50-56
HASQSIS
L2
of SEQ ID
NO.: 25
AE11-5 VL
CDR-
Residues 89-98
HQSSSSPPPT
L3
of SEQ ID
NO.: 25
TNF-JK1 VH
26
EVQLVESGGGLVQPGGSLRLSCATSGFTFN
NYWMSWVRQAPGKGLEWVANINHDESEKYY
VDSAKGRFTISRDNAEKSLFLQMNSLRAED
TAVYYCARIIRGRVGFDYYNYAMDVWGQGT
LVTVSS
TNF-JK1 VH
CDR-
Residues 31-35
NYWMS
H1
of SEQ ID
NO.: 26
TNF-JK1 VH
CDR-
Residues 50-66
NINHDESEKYYVDSAKG
H2
of SEQ ID
NO.: 26
TNF-JK1 VH
CDR-
Residues 99-115
IIRGRVGFDYYNYAMDV
H3
of SEQ ID
NO.: 26
TNF-JK1 VL
27
DIRLTQSPSPLSASVGDRVTITCRASQSIG
NYLNWYQHKPGKAPKLLIYAASSLQSGVPS
RFSGTGSGTDFTLTISSLQPEDFATYYCQE
SYSLIFAGGTKVEIK
TNF-JK1 VL
CDR-
Residues 24-34
RASQSIGNYLN
L1
of SEQ ID
NO.: 27
TNF-JK1 VL
CDR-
Residues 50-56
AASSLQS
L2
of SEQ ID
NO.: 27
TNF-JK1 VL
CDR-
Residues 89-95
QESYSLI
L3
of SEQ ID
NO.: 27
TNF-Y7C VH
28
EVQLVQSGAEVKKPGASVKVSCKTSGYTFS
NYDINWVRQPTGQGLEWMGWMDPNNGNTGY
AQKFVGRVTMTRDTSKTTAYLELSGLKSED
TAVYYCARSSGSGGTWYKEYFQSWGQGTMV
TVSS
TNF-Y7C VH
CDR-
Residues 31-35
NYDIN
H1
of SEQ ID
NO.: 28
TNF-Y7C VH
CDR-
Residues 50-66
WMDPNNGNTGYAQKFVG
H2
of SEQ ID
NO.: 28
TNF-Y7C VH
CDR-
Residues 99-112
KSSGSGGTWYKEYFQS
H3
of SEQ ID
NO.: 28
TNF-Y7C VL
29
DIVMTQSPLSLPVTPGEPASISCRSSQSLL
HSNGYNYLDWYLQKPGQFPQLLIYLGSYRA
SGVPDRFSGSGSGTDFTLKISRVEAEDVGV
YYCMQRIEFPPGTFGQGTKLGIK
TNF-Y7C VL
CDR-
Residues 24-39
RSSQSLLHSNGYNYLD
L1
of SEQ ID
NO.: 29
TNF-Y7C VL
CDR-
Residues 55-61
LGSYRAS
L2
of SEQ ID
NO.: 29
TNF-Y7C VL
CDR-
Residues 94-103
MQRIEFPPGT
L3
of SEQ ID
NO.: 29
AE11-7 VH
30
EVQLVQSGAEVKKPGASVKVSCKTSGYSLT
QYPIHWVRQAPGQRPEWMGWISPGNGNTKL
SPKFQGRVTLSRDASAGTVFMDLSGLTSDD
TAVYFCTSVDLGDHWGQGTLVTVSS
AE11-7 VH
CDR-
Residues 31-35
QYPIH
H1
of SEQ ID
NO.: 30
AE11-7 VH
CDR-
Residues 50-66
WISPGNGNTKLSPKFQG
H2
of SEQ ID
NO.: 30
AE11-7 VH
CDR-
Residues 99-104
VDLGDH
H3
of SEQ ID
NO.: 30
AE11-7 VL
31
DIVMTQSPEFQSVTPKEKVTITCRASQSIG
SSLHWYQQKPDQSPKLLINYASQSFSGVPS
RFSGGGSGTDFTLTINSLEAEDAATYYCHQ
SSNLPITFGQGTRLEIK
AE11-7 VL
CDR-
Residues 24-34
RASQSIGSSLH
L1
of SEQ ID
NO.: 31
AE11-7 VL
CDR-
Residues 50-56
YASQSFS
L2
of SEQ ID
NO.: 31
AE11-7 VL
CDR-
Residues 89-97
HQSSNLPIT
L3
of SEQ ID
NO.: 31
AE11-13 VH
32
EVQLVESGGGLVQPGRSLRLSCAASGFTFD
DYPMHWVRQAPGEGLEWVSGISSNSASIGY
ADSVKGRFTISRDNAQNTLYLQMNSLGDED
TAVYYCVSLTLGIGQGTLVTVSS
AE11-13 VH
CDR-
Residues 31-35
DYPMH
H1
of SEQ ID
NO.: 32
AE11-13 VH
CDR-
Residues 50-66
GISSNSASIGYADSVKG
H2
of SEQ ID
NO.: 32
AE11-13 VH
CDR-
Residues 99-102
LTLG
H3
of SEQ ID
NO.: 32
AE11-13 VL
33
DIRLTQSPSSLSASVGDRVTITCRASQSIG
NYLHWYQQKPGKAPKLLIYAASSLQSGVPS
RFSGSGSGTDFTLTISSLQPEDFATYYCQQ
SYSTLYSFGQGTKLEIK
AE11-13 VL
CDR-
Residues 24-34
RASQSIGNYLH
L1
of SEQ ID
NO.: 33
AE11-13 VL
CDR-
Residues 50-56
AASSLQS
L2
of SEQ ID
NO.: 33
AE11-13 VL
CDR-
Residues 89-97
QQSYSTLYS
L3
of SEQ ID
NO.: 33
The AE11-5 human antibody to human TNF was affinity matured by in vitro display technology. One light chain library was constructed to contain limited mutagenesis at the following residues: 28, 31, 32, 51, 55, 91, 92, 93, 95a and 96 (Kabat numbering). This library also contained framework germline back-mutations D1E, M4L, H11Q, R49K, H76N and Q103K as well as toggled residues at position 50(R/K) and 94(S/L) to allow for framework germlining during library selections. Two heavy chain libraries were made to contain limited mutagenesis in CDRH1 and CDRH2 at residues 30, 31, 33, 50, 52, and 55 to 58 (Kabat numbering) or in CDRH3 at residues 95 to 100b. The library containing CDRH1 and CDRH2 diversities also had framework germline back-mutations Al8V and L64Q and toggled residue at 54(L/F) and 78(V/A). The CDRH3 library has an additional toggled residue at 100c(A/F).
All three libraries were selected separately for the ability to bind human or cynomolgus monkey TNF in the presence of decreasing concentrations of biotinylated human or cynomolgus monkey TNF antigens. All mutated CDR sequences recovered from library selections were recombined into additional libraries and the recombined libraries were subjected to more stringent selection conditions before individual antibodies are identified.
Table 11 provides a list of amino acid sequences of VH regions of affinity matured fully human TNF antibodies derived from AE11-5. Amino acid residues of individual CDRs of each VH sequence are indicated in bold.
TABLE 11
List of amino acid sequences of affinity
matured AE11-5 VH variants
Clone
SEQ ID NO:
VH
J685M2S2-10VH
94
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSWYA
ISWVRQAPGQGLEWMGGITPILGSANYAQKFQG
RVTITADESTSTAYMELSSLRSEDTAVYYCARG
LYYDPTRADYWGQGTLVTVSS
J685M2S2-12VH
95
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSYYS
ISWVRQAPGQGLEWMGGIMPILGTANYAQKFQG
RVTITADESTSTAYMELSSLRSEDTAVYYCARG
LYYDPTRADYWGQGTLVTVSS
J685M2S2-13VH
96
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSWYA
ISWVRQAPGQGLEWMGGIIPILGSPIYAQKFQG
RVTITADESTSTAYMELSSLRSEDTAVYYCARG
LYYDPTRADYWGQGTLVTVSS
J685M2S2-14VH
97
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSWYT
ISWVRQAPGQGLEWMGGIIPILGSPIYAQKFQG
RVTITADESTSTAYMELSSLRSEDTAVYYCARG
LYYDPTRADYWGQGTLVTVSS
J685M2S2-16VH
98
EVQLVQSGAEVKKPGSSVKVSCKASGGTFAWYS
ISWVRQAPGQGLEWMGGITPILGTANYAQKFQG
RVTITADESTSTAYMELSSLRSEDTAVYYCARG
LYYDPTRADYWGQGTLVTVSS
J685M2S2-18VH
99
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSFYA
ISWVRQAPGQGLEWMGGITPILGAATYAQKFQG
RVTITADESTSTAYMELSSLRSEDTAVYYCARG
LYYDPTRADYWGQGTLVTVSS
J685M2S2-1VH
100
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSIYA
ISWVRQAPGQGLEWMGGITPILGAAVYAQKFQG
RVTITADESTSTAYMELSSLRSEDTAVYYCARG
LYYDPTRADYWGQGTLVTVSS
J685M2S2-21VH
101
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSWYT
ISWVRQAPGQGLEWMGGIMPILGTANYAQKFQG
RVTITADESTSTAYMELSSLRSEDTAVYYCARG
LYYDPTRADYWGQGTLVTVSS
J685M2S2-23VH
102
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSWYA
ISWVRQAPGQGLEWMGGITPILGVAVYAQKFQG
RVTITADESTSTAYMELSSLRSEDTAVYYCARG
LYYDPTRADYWGQGTLVTVSS
J685M2S2-25VH
103
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSWYA
ISWVRQAPGQGLEWMGGITPILGTANYAQKFQG
RVTITADESTSTAYMELSSLRSEDTAVYYCARG
LYYDPTRADYWGQGTLVTVSS
J685M2S2-27VH
104
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSWYA
ISWVRQAPGQGLEWMGGITPILGSAHYAQKFQG
RVTITADESTSTAYMELSSLRSEDTAVYYCARG
LYYDPTRADYWGQGTLVTVSS
J685M2S2-28VH
105
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSWYA
ISWVRQAPGQGLEWMGGITPILGSAIYAQKFQG
RVTITADESTSTVYMELSSLRSEDTAVYYCARG
LYYDPTRADYWGQGTLVTVSS
J685M2S2-29VH
106
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSWYA
ISWVRQAPGQGLEWMGGITPILGTAIYAQKFQG
RVTITADESTSTVYMELSSLRSEDTAVYYCARG
LYYDPTRADYWGQGTLVTVSS
J685M2S2-31VH
107
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSIYT
ISWVRQAPGQGLEWMGGIIPILRNPIYAQKFQG
RVTITADESTSTAYMELSSLRSEDTAVYYCARG
LYYDPTRADYWGQGTLVTVSS
J685M2S2-32VH
108
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSYYA
ISWVRQAPGQGLEWMGGIMPILGTPTYAQKFQG
RVTITADESTSTAYMELSSLRSEDTAVYYCARG
LYYDPTRADYWGQGTLVTVSS
J685M2S2-35VH
109
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSIYT
ISWVRQAPGQGLEWMGGIIPILGAPIYAQKFQG
RVTITADESTSTAYMELSSLRSEDTAVYYCARG
LYYDPTRADYWGQGTLVTVSS
J685M2S2-37VH
110
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSWYA
ISWVRQAPGQGLEWMGGITPILGSATYAQKFQG
RVTITADESTSTAYMELSSLRSEDTAVYYCARG
LYYDPTRADYWGQGTLVTVSS
J685M2S2-38VH
111
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSWYS
ISWVRQAPGQGLEWMGGIMPILGSASYAQKFQG
RVTITADESTSTAYMELSSLRSEDTAVYYCARG
LYYDPTRADYWGQGTLVTVSS
J685M2S2-43VH
112
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSWYT
ISWVRQAPGQGLEWMGGIMPILGTASYAQKFQG
RVTITADESTSTAYMELSSLRSEDTAVYYCARG
LYYDPTRADYWGQGTLVTVSS
J685M2S2-44VH
113
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSWYS
ISWVRQAPGQGLEWMGGITPILGTANYAQKFQG
RVTITADESTSTAYMELSSLRSEDTAVYYCARG
LYYDPTRADYWGQGTLVTVSS
J685M2S2-45VH
114
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSWYA
ISWVRQAPGQGLEWMGGIMPILGTATYAQKFQG
RVTITADESTSTAYMELSSLRSEDTAVYYCARG
LYYDPTRADYWGQGTLVTVSS
J685M2S2-46VH
115
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSFYT
ISWVRQAPGQGLEWMGGIMPILGSPHYAQKFQG
RVTITADESTSTAYMELSSLRSEDTAVYYCARG
LYYDPTRADYWGQGTLVTVSS
J685M2S2-47VH
116
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSWYA
ISWVRQAPGQGLEWMGGITPILGTANYAQKFQG
RVTITADESTSTVYMELSSLRSEDTAVYYCARG
LYYDPTRADYWGQGTLVTVSS
J685M2S2-48VH
117
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSWYA
ISWVRQAPGQGLEWMGGIMPILGSATYAQKFQG
RVTITADESTSTAYMELSSLRSEDTAVYYCARG
LYYDPTRADYWGQGTLVTVSS
J685M2S2-4VH
118
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSWYA
ISWVRQAPGQGLEWMGGIIPILGTPTYAQKFQG
RVTITADESTSTAYMELSSLRSEDTAVYYCARG
LYYDPTRADYWGQGTLVTVSS
J685M2S2-50VH
119
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYA
ISWVRQAPGQGLEWMGGIIPILGTANYAQKFQG
RVTITADESTSTVYMELSSLRSEDTAVYYCARG
VYYDPKRADYWGQGTLVTVSS
J685M2S2-51VH
120
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSLYT
ISWVRQAPGQGLEWMGGIMPILGAPRYAQKFQG
RVTITADESTSTAYMELSSLRSEDTAVYYCARG
LYYDPTRADYWGQGTLVTVSS
J685M2S2-52VH
121
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSYYA
ISWVRQAPGQGLEWMGGIMPILGSPIYAQKFQG
RVTITADESTSTVYMELSSLRSEDTAVYYCARG
LYYDPTRADYWGQGTLVTVSS
J685M2S2-53VH
122
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSIYA
ISWVRQAPGQGLEWMGGILPILGSPIYAQKFQG
RVTITADESTSTAYMELSSLRSEDTAVYYCARG
LYYDPTRADYWGQGTLVTVSS
J685M2S2-55VH
123
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSIYA
ISWVRQAPGQGLEWMGGIIPILGSPIYAQKFQG
RVTITADESTSTAYMELSSLRSEDTAVYYCARG
LYYDPTRADYWGQGTLVTVSS
J685M2S2-56VH
124
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSWYA
ISWVRQAPGQGLEWMGGIVPILGAPLYAQKFQG
RVTITADESTSTAYMELSSLRSEDTAVYYCARG
LYYDPTRADYWGQGTLVTVSS
J685M2S2-58VH
125
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSIYA
ISWVRQAPGQGLEWMGGIMPILGAPIYAQKFQG
RVTITADESTSTAYMELSSLRSEDTAVYYCARG
LYYDPTRADYWGQGTLVTVSS
J685M2S2-5VH
126
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSYYT
ISWVRQAPGQGLEWMGGIMPILGTPAYAQKFQG
RVTITADESTSTAYMELSSLRSEDTAVYYCARG
LYYDPTRADYWGQGTLVTVSS
J685M2S2-61VH
127
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSWYS
ISWVRQAPGQGLEWMGGITPILGAATYAQKFQG
RVTITADESTSTAYMELSSLRSEDTAVYYCARG
LYYDPTRADYWGQGTLVTVSS
J685M2S2-62VH
128
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSWYA
ISWVRQAPGQGLEWMGGIIPILGTPTYAQKFQG
RVTITADESTSTVYMELSSLRSEDTAVYYCARG
LYYDPTRADYWGQGTLVTVSS
J685M2S2-63VH
129
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSWYA
ISWVRQAPGQGLEWMGGIIPILGTPIYAQKFQG
RVTITADESTSTAYMELSSLRSEDTAVYYCARG
LYYDPTRADYWGQGTLVTVSS
J685M2S2-64VH
130
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSWYA
ISWVRQAPGQGLEWMGGITPILGIGNYAQKFQG
RVTITADESTSTAYMELSSLRSEDTAVYYCARG
LYYDPTRADYWGQGTLVTVSS
J685M2S2-66VH
131
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSIYA
ISWVRQAPGQGLEWMGGIVPILGAATYAQKFQG
RVTITADESTSTAYMELSSLRSEDTAVYYCARG
LYYDPTRADYWGQGTLVTVSS
J685M2S2-67VH
132
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSWYA
ISWVRQAPGQGLEWMGGITPILGSSTYAQKFQG
RVTITADESTSTAYMELSSLRSEDTAVYYCARG
LYYDPTRADYWGQGTLVTVSS
J685M2S2-68VH
133
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSWYT
ISWVRQAPGQGLEWMGGIMPILGTANYAQKFQG
RVTITADESTSTAYMELSSLRSEDTAVYYCARG
LYYDPTRADYWGQGTLVTVSS
J685M2S2-6VH
134
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSWYA
ISWVRQAPGQGLEWMGGITPILGNSIYAQKFQG
RVTITADESTSTAYMELSSLRSEDTAVYYCARG
LYYDPTRADYWGQGTLVTVSS
J685M2S2-70VH
135
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSWYA
ISWVRQAPGQGLEWMGGITPILGSPIYAQKFQG
RVTITADESTSTAYMELSSLRSEDTAVYYCARG
LYYDPTRADYWGQGTLVTVSS
J685M2S2-71VH
136
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSWYA
ISWVRQAPGQGLEWMGGIMPILGTPTYAQKFQG
RVTITADESTSTAYMELSSLRSEDTAVYYCARG
LYYDPTRADYWGQGTLVTVSS
J685M2S2-72VH
137
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSYYA
ISWVRQAPGQGLEWMGGITPILGAANYAQKFQG
RVTITADESTSTVYMELSSLRSEDTAVYYCARG
LYYDPTRADYWGQGTLVTVSS
J685M2S2-73VH
138
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSWYA
ISWVRQAPGQGLEWMGGITPILGAAIYAQKFQG
RVTITADESTSTAYMELSSLRSEDTAVYYCARG
LYYDPTRADYWGQGTLVTVSS
J685M2S2-75VH
139
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSWYA
ISWVRQAPGQGLEWMGGITPILGTATYAQKFQG
RVTITADESTSTAYMELSSLRSEDTAVYYCARG
LYYDPTRADYWGQGTLVTVSS
J685M2S2-76VH
140
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSWYS
ISWVRQAPGQGLEWMGGITPILGSAHYAQKFQG
RVTITADESTSTAYMELSSLRSEDTAVYYCARG
LYYDPTRADYWGQGTLVTVSS
J685M2S2-77VH
141
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSWYA
ISWVRQAPGQGLEWMGGITPILGNAIYAQKFQG
RVTITADESTSTAYMELSSLRSEDTAVYYCARG
LYYDPTRADYWGQGTLVTVSS
J685M2S2-78VH
142
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSWYA
ISWVRQAPGQGLEWMGGITPILRSAVYAQKFQG
RVTITADESTSTVYMELSSLRSEDTAVYYCARG
LYYDPTRADYWGQGTLVTVSS
J685M2S2-7VH
143
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSWYS
ISWVRQAPGQGLEWMGGIMPILGTANYAQKFQG
RVTITADESTSTAYMELSSLRSEDTAVYYCARG
LYYDPTRADYWGQGTLVTVSS
J685M2S2-80VH
144
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSWYA
ISWVRQAPGQGLEWMGGITPILGTASYAQKFQG
RVTITADESTSTAYMELSSLRSEDTAVYYCARG
LYYDPTRADYWGQGTLVTVSS
J685M2S2-81VH
145
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSWYA
ISWVRQAPGQGLEWMGGITPILGTAIYAQKFQG
RVTITADESTSTAYMELSSLRSEDTAVYYCARG
LYYDPTRADYWGQGTLVTVSS
J685M2S2-82VH
146
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSWYA
ISWVRQAPGQGLEWMGGITPILGSPAYAQKFQG
RVTITADESTSTAYMELSSLRSEDTAVYYCARG
LYYDPTRADYWGQGTLVTVSS
J685M2S2-83VH
147
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSRYA
ISWVRQAPGQGLEWMGGIIPILGPASYAQKFQG
RVTITADESTSTAYMELSSLRSEDTAVYYCARG
LYYDPTRADYWGQGTLVTVSS
J685M2S2-84VH
148
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSWYA
ISWVRQAPGQGLEWMGGITPILDAAIYAQKFQG
RVTITADESTSTAYMELSSLRSEDTAVYYCARG
LYYDPTRADYWGQGTLVTVSS
J685M2S2-86VH
149
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSWYT
ISWVRQAPGQGLEWMGGIMPILGIPNYAQKFQG
RVTITADESTSTAYMELSSLRSEDTAVYYCARG
LYYDPTRADYWGQGTLVTVSS
J685M2S2-87VH
150
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSIYA
ISWVRQAPGQGLEWMGGITPILGSAIYAQKFQG
RVTITADESTSTAYMELSSLRSEDTAVYYCARG
LYYDPTRADYWGQGTLVTVSS
J685M2S2-88VH
151
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSYYA
ISWVRQAPGQGLEWMGGIMPILGTATYAQKFQG
RVTITADESTSTVYMELSSLRSEDTAVYYCARG
LYYDPTRADYWGQGTLVTVSS
J685M2S2-89VH
152
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYA
ISWVRQAPGQGLEWMGGIIPILGTANYAQKFQG
RVTITADESTSTVYMELSSLRSEDTAVYYCARG
VYFDPKRADYWGQGTLVTVSS
J685M2S2-8VH
153
EVQLVQSGAEVKKPGSSVKVSCKASGGTFNWYT
ISWVRQAPGQGLEWMGGIMPILGTANYAQKFQG
RVTITADESTSTAYMELSSLRSEDTAVYYCARG
LYYDPTRADYWGQGTLVTVSS
J685M2S2-90VH
154
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYA
ISWVRQAPGQGLEWMGGIIPILGTANYAQKFQG
RVTITADESTSTVYMELSSLRSEDTAVYYCARG
VYFDFTRADYWGQGTLVTVSS
J685M2S2-91VH
155
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSWYA
ISWVRQAPGQGLEWMGGIIPILRFPTYAQKFQG
RVTITADESTSTVYMELSSLRSEDTAVYYCARG
LYYDPTRADYWGQGTLVTVSS
J685M2S2-92VH
156
EVQLVQSGAEVKKPGSSVKVSCKVSGGTFSWYS
ISWVRQAPGQGLEWMGGILPILDTANYAQKFQG
RVTITADESTSTAYMELSSLRSEDTAVYYCARG
LYYDPTRADYWGQGTLVTVSS
J685M2S2-93VH
157
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSWYA
ISWVRQAPGQGLEWMGGIMPILGTAVYAQKFQG
RVTITADESTSTAYMELSSLRSEDTAVYYCARG
LYYDPTRADYWGQGTLVTVSS
J685M2S2-94VH
158
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSIYS
ISWVRQAPGQGLEWMGGILPILGTANYAQKFQG
RVTITADESTSTVYMELSSLRSEDTAVYYCARG
LYYDPTRADYWGQGTLVTVSS
J688M2-11VH
159
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYA
ISWVRQAPGQGLEWMGGIIPILGTANYAQKFQG
RVTITADESTSTVYMELSSLRSEDTAVYYCARG
VYYDPTRADYWGQGTLVTVSS
J688M2-13VH
160
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYA
ISWVRQAPGQGLEWMGGIIPILGTANYAQKFQG
RVTITADESTSTVYMELSSLRSEDTAVYYCARG
LYYDPSRADYWGQGTLVTVSS
J688M2-14VH
161
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYA
ISWVRQAPGQGLEWMGGIIPILGTANYAQKFQG
RVTITADESTSTVYMELSSLRSEDTAVYYCARG
VYFNPTRADYWGQGTLVTVSS
J688M2-16VH
162
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYA
ISWVRQAPGQGLEWMGGIIPILGTANYAQKFQG
RVTITADESTSTVYMELSSLRSEDTAVYYCARG
LYYDPARFDYWGQGTLVTVSS
J688M2-20VH
163
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYA
ISWVRQAPGQGLEWMGGIIPILGTANYAQKFQG
RVTITADESTSTVYMELSSLRSEDTAVYYCARG
LYYNPSRADYWGQGTLVTVSS
J688M2-21VH
164
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYA
ISWVRQAPGQGLEWMGGIIPILGTANYAQKFQG
RVTITADESTSTVYMELSSLRSEDTAVYYCARG
LYYDPKRADYWGQGTLVTVSS
J688M2-22VH
165
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYA
ISWVRQAPGQGLEWMGGIIPILGTANYAQKFQG
RVTITADESTSTVYMELSSLRSEDTAVYYCARG
LYYDPRRADYWGQGTLVTVSS
J688M2-28VH
166
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYA
ISWVRQAPGQGLEWMGGIIPILGTANYAQKFQG
RVTITADESTSTVYMELSSLRSEDTAVYYCARG
FYYDPTRADYWGQGTLVTVSS
J688M2-29VH
167
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYA
ISWVRQAPGQGLEWMGGIIPILGTANYAQKFQG
RVTITADESTSTVYMELSSLRSEDTAVYYCARG
LYYDFTRADYWGQGTLVTVSS
J688M2-2VH
168
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYA
ISWVRQAPGQGLEWMGGIIPILGTANYAQKFQG
RVTITADESTSTVYMELSSLRSEDTAVYYCARG
LYFDPKRADYWGQGTLVTVSS
J688M2-37VH
169
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYA
ISWVRQAPGQGLEWMGGIIPILGTANYAQKFQG
RVTITADESTSTVYMELSSLRSEDTAVYYCARG
VYFDPTRADYWGQGTLVTVSS
J688M2-3VH
170
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYA
ISWVRQAPGQGLEWMGGIIPILGTANYAQKFQG
RVTITADESTSTVYMELSSLRSEDTAVYYCARG
IYYDPSRADYWGQGTLVTVSS
J688M2-46VH
171
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYA
ISWVRQAPGQGLEWMGGIIPILGTANYAQKFQG
RVTITADESTSTVYMELSSLRSEDTAVYYCARS
LYYERTRADYWGQGTLVTVSS
J688M2-48VH
172
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYA
ISWVRQAPGQGLEWMGGIIPILGTANYAQKFQG
RVTITADESTSTVYMELSSLRSEDTAVYYCARW
RFYIPIRFDYWGQGTLVTVSS
J688M2-4VH
173
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYA
ISWVRQAPGQGLEWMGGIIPILGTANYAQKFQG
RVTITADESTSTVYMELSSLRSEDTAVYYCARG
VYYDFTRADYWGQGTLVTVSS
J688M2-50VH
174
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYA
ISWVRQAPGQGLEWMGGIIPILGTANYAQKFQG
RVTITADESTSTVYMELSSLRSEDTAVYYCARG
LFYDPSRADYWGQGTLVTVSS
J688M2-52VH
175
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYA
ISWVRQAPGQGLEWMGGIIPILGTANYAQKFQG
RVTITADESTSTVYMELSSLRSEDTAVYYCARG
VYYDPVRADYWGQGTLVTVSS
J688M2-56VH
176
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYA
ISWVRQAPGQGLEWMGGIIPILGTANYAQKFQG
RVTITADESTSTVYMELSSLRSEDTAVYYCARG
VYYDPIRADYWGQGTLVTVSS
J688M2-57VH
177
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYA
ISWVRQAPGQGLEWMGGIIPILGTANYAQKFQG
RVTITADESTSTVYMELSSLRSEDTAVYYCARG
IYYDPKRADYWGQGTLVTVSS
J688M2-58VH
178
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYA
ISWVRQAPGQGLEWMGGIIPILGTANYAQKFQG
RVTITADESTSTVYMELSSLRSEDTAVYYCARG
LYYNPIRFDYWGQGTLVTVSS
J688M2-64VH
179
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYA
ISWVRQAPGQGLEWMGGIIPILGTANYAQKFQG
RVTITADESTSTVYMELSSLRSEDTAVYYCARG
VYFDPARADYWGQGTLVTVSS
J688M2-65VH
180
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYA
ISWVRQAPGQGLEWMGGIIPILGTANYAQKFQG
RVTITADESTSTVYMELSSLRSEDTAVYYCARG
VFFDPTRADYWGQGTLVTVSS
J688M2-68VH
181
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYA
ISWVRQAPGQGLEWMGGIIPILGTANYAQKFQG
RVTITADESTSTVYMELSSLRSEDTAVYYCARG
VFYNPTRADYWGQGTLVTVSS
J688M2-69VH
182
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYA
ISWVRQAPGQGLEWMGGIIPILGTANYAQKFQG
RVTITADESTSTVYMELSSLRSEDTAVYYCARG
LYYEGPSADYWGQGTLVTVSS
J688M2-6VH
183
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYA
ISWVRQAPGQGLEWMGGIIPILGTANYAQKFQG
RVTITADESTSTVYMELSSLRSEDTAVYYCARG
LYYAPNRADYWGQGTLVTVSS
J688M2-73VH
184
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYA
ISWVRQAPGQGLEWMGGIIPILGTANYAQKFQG
RVTITADESTSTVYMELSSLRSEDTAVYYCARG
LFYDPTRADYWGQGTLVTVSS
J688M2-74VH
185
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYA
ISWVRQAPGQGLEWMGGIIPILGTANYAQKFQG
RVTITADESTSTVYMELSSLRSEDTAVYYCARG
VYYNPTRADYWGQGTLVTVSS
J688M2-75VH
186
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYA
ISWVRQAPGQGLEWMGGIIPILGTANYAQKFQG
RVTITADESTSTVYMELSSLRSEDTAVYYCARG
VYYDPARADYWGQGTLVTVSS
J688M2-7VH
187
EVQLVQSGAEVKKSGSSVKVSCKASGGTFSSYA
ISWVRQAPGQGLEWMGGIIPILGTANYAQKFQG
RVTITADESTSTVYMELSSLRSEDTAVYYCARG
VYYDPGRADYWGQGTLVTVSS
J688M2-81VH
188
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYA
ISWVRQAPGQGLEWMGGIIPILGTANYAQKFQG
RVTITADESTSTVYMELSSLRSEDTAVYYCARG
VYFDPSRADYWGQGTLVTVSS
J688M2-82VH
189
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYA
ISWVRQAPGQGLEWMGGIIPILGTANYAQKFQG
RVTITADESTSTVYMELSSLRSEDTAVYYCARG
LYFDPSRFDYWGQGTLVTVSS
J688M2-83VH
190
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYA
ISWVRQAPGQGLEWMGGIIPILGTANYAQKFQG
RVTITADESTSTVYMELSSLRSEDTAVYYCARG
LYFDFTRADYWGQGTLVTVSS
J688M2-84VH
191
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYA
ISWVRQAPGQGLEWMGGIIPILGTANYAQKFQG
RVTITADESTSTVYMELSSLRSEDTAVYYCARG
IYYDPTRADYWGQGTLVTVSS
J688M2-88VH
192
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYA
ISWVRQAPGQGLEWMGGIIPILGTANYAQKFQG
RVTITADESTSTVYMELSSLRSEDTAVYYCARG
LYFDPSRADYWGQGTLVTVSS
J688M2-89VH
193
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYA
ISWVRQAPGQGLEWMGGIIPILGTANYAQKFQG
RVTITADESTSTVYMELSSLRSEDTAVYYCARG
VYYDPSRFDYWGQGTLVTVSS
J688M2-8VH
194
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYA
ISWVRQAPGQGLEWMGGIIPILGTANYAQKFQG
RVTITADESTSTVYMELSSLRSEDTAVYYCARG
QYYDTSRADYWGQGTLVTVSS
J688M2-90VH
195
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYA
ISWVRQAPGQGLEWMGGIIPILGTANYAQKFQG
RVTITADESTSTVYMELSSLRSEDTAVYYCARS
LYYDTTRFDYWGQGTLVTVSS
J688M2-92VH
196
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYA
ISWVRQAPGQGLEWMGGIIPILGTANYAQKFQG
RVTITADESTSTVYMELSSLRSEDTAVYYCARG
VFYDPTRADYWGQGTLVTVSS
J688M2-94VH
197
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYA
ISWVRQAPGQGLEWMGGIIPILGTANYAQKFQG
RVTITADESTSTVYMELSSLRSEDTAVYYCARG
IYYDPARADYWGQGTLVTVSS
J688M2-95VH
198
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYA
ISWVRQAPGQGLEWMGGIIPILGTANYAQKFQG
RVTITADESTSTVYMELSSLRSEDTAVYYCARG
LFYDPRRADYWGQGTLVTVSS
J688M2-96VH
199
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYA
ISWVRQAPGQGLEWMGGIIPILGTANYAQKFQG
RVTITADESTSTVYMELSSLRSEDTAVYYCARG
VYYDTTRADYWGQGTLVTVSS
J693FRM2S2L-
200
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYA
32VH
ISWVRQAPGQGLEWMGGIIPILGTANYAQKFQG
RVTITADESTSTVYMELSSLRSEDTAVYYCARG
LYYDPARADYWGQGTLVTVSS
J693FRM2S2L-
201
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYA
40VH
ISWVRQAPGQGLEWMGGIIPILGTANYAQKFQG
RVTITADESTSTVYMELSSLRSEDTAVYYCSRG
LYYDPTRADYWGQGTLVTVSS
J693FRM2S2L-
202
EVQLVQSGAEVMKPGSSVKVSCKASGGTFSSYA
70VH
ISWVRQAPGQGLEWMGGIIPILGTANYAQKFQG
RVTITADESTSTVYMELSSLRSEDTAVYYCTRG
LYYDPTRADYWGQGTLVTVSS
J693FRM2S2R-
203
EVQLVQSGAEVKKPGASVKVSCKASGYTFNNYA
29VH
ISWVRQAPGQGLEWMGGIIPILGTANYAQKFQG
RVTITADESTSTVYMELSSLRSEDTAVYYCARG
LYYDPTRADYWGQGTLVTVSS
J693FRM2S2R-
204
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYA
46VH
ISWVRQAPGQGLEWMGGIIPILGTANYAQKFQG
RVTITADESTSTVYMELSSLRSEDTAVYYCTRG
LYYDPTRADYWGQGTLVTVSS
J693FRM2S2R-
205
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYA
65VH
ISWVRQAPGQGLEWMGGIIPILGTANYAQKFQG
RVTITADESTSTVYMELSSLRSEDTAVYYCTRG
IYYDPTRADYWGQGTLVTVSS
J693M2S2L-17VH
206
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYA
ISWVRQAPGQGLEWMGGIIPILGTANYAQEFQG
RVTITADESTSTVYMELSSLRSEDTAVYYCARG
LYYDPTRADYWGQGTLVTVSS
J693M2S2L-32VH
207
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYA
ISWVRQAPGQGLEWMGGIIPILGTANYAQKFQG
RVTITADESTSTVYMELSSLRSEDTAVYYCVRG
LYYDPTRADYWGQGTLVTVSS
J693M2S2L-67VH
208
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYA
ISWVRQAPGQGLEWMGGIIPILGTASYAQKFQG
RVTITADESTSTVYMELSSLRSEDTAVYYCARG
LYYDPTRADYWGQGTLVTVSS
J693M2S2L-75VH
209
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYA
ISWVRQAPGQGLEWMGGIIPILGTANYAQKFQG
RVTITADESTSTVYMELSSLRSEDTAVYYCAKG
LYYDPTRADYWGQGTLVTVSS
J693M2S2L-78VH
210
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYA
ISWVRQAPGQGLEWMGGIIPILGTANYAQKFQG
RVTITADESTSTVYMELSSLRSEDTAVYYCERG
LYYDPTRADYWGQGTLVTVSS
J693M2S2L-79VH
211
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSNYA
ISWVRQAPGQGLEWMGGIIPILGTANYAQKFQG
RVTITADESTSTVYMELSSLRSEDTAVYYCARG
LYYDPTRADYWGQGTLVTVSS
J693M2S2L-94VH
212
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYA
ISWVRQAPGQGLEWMGGIIPILGTANYAHKFQG
RVTITADESTSTVYMELSSLRSEDTAVYYCARG
LYYDPTRADYWGQGTLVTVSS
J693M2S2R-22VH
213
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYA
ISWVRQAPGQGLEWMGGIIPILGTANYAQKFQG
RVTITADESTSTVYMELSSLRSEDTAVYYCARG
LYYDPTRADCWGQGTLVTVSS
J693M2S2R-24VH
214
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYA
ISWVQQAPGQGLEWMGGIIPILGTANYAQKFQG
RVTITADESTSTVYMELSSLRSEDTAVYYCARG
LYYDPTRADYWGQGTLVTVSS
J693M2S2R-2VH
215
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYA
ISWVRQAPGQGLEWMGGITPILGTANYAQKFQG
RVTITADESTSTVYMELSSLRSEDTAVYYCARG
LYYDPTRADYWGQGTLVTVSS
J693M2S2R-31VH
216
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYA
ISWVRQAPGQGLEWMGGIIPILGTANYAQKFQG
RVTITADESTSTVYMELSSLRSEDTAVYYCARG
LYYDPTRADYWGQGTLVTVSS
J693M2S2R-71VH
217
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYA
TSWVRQAPGQGLEWMGGIIPILGTANYAQKFQG
RVTITADESTSTVYMELSSLRSEDTAVYYCARG
LYYDPTRADYWGQGTLVTVSS
J693M2S2R-84VH
218
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYA
ISWVRQAPGQGLEWMGGIIPILGTANYAQKFLG
RVTITADESTSTVYMELSSLRSEDTAVYYCARG
LYYDPTRADYWGQGTLVTVSS
J693M2S2R-89VH
219
EVQLVQSGAEVKKPGSSVKVSCKASGGTSSSYA
ISWVRQAPGQGLEWMGGIIPILGTANYAQKFQG
RVTITADESTSTVYMELSSLRSEDTAVYYCARG
LYYDPTRADYWGQGTLVTVSS
J703M1S3-10VH
220
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSWYA
ISWVRQAPGQGLEWMGGITPILGSATYAQKFQG
RVTITADESTSTVYMELSSLRSEDTAVYYCARG
LYYDPKRADYWGQGTLVTVSS
J703M1S3-11VH
221
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSWYA
ISWVRQAPGQGLEWMGGITPILGAASYAQKFQG
RVTITADESTSTVYMELSSLRSEDTAVYYCARG
IYYDPTRADYWGQGTLVTVSS
J703M1S3-12VH
222
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSWYA
ISWVRQAPGQGLEWMGGITPILGAASYAQKFQG
RVTITADESTSTVYMELSSLRSEDTAVYYCARG
IYYDPARADYWGQGTLVTVSS
J703M1S3-13VH
223
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSWYA
ISWVRQAPGQGLEWMGGITPILGAANYAQKFQG
RVTITADESTSTVYMELSSLRSEDTAVYYCARG
VYYDPKRADYWGQGTLVTVSS
J703M1S3-14VH
224
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSWYT
ISWVRQAPGQGLEWMGGIMPILGSPTYAQKFQG
RVTITADESTSTVYMELSSLRSEDTAVYYCARG
VYYDPRRADYWGQGTLVTVSS
J703M1S3-16VH
225
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSWYA
ISWVRQAPGQGLEWMGGITPILGSATYAQKFQG
RVTITADESTSTVYMELSSLRSEDTAVYYCARG
IYYDPKRADYWGQGTLVTVSS
J703M1S3-17VH
226
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSWYA
ISWVRQAPGQGLEWMGGIVPILGTPIYAQKFQG
RVTITADESTSTVYMELSSLRSEDTAVYYCARG
VYYDPKRADYWGQGTLVTVSS
J703M1S3-18VH
227
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSWYA
ISWVRQAPGQGLEWMGGITPILGSANYAQKFQG
RVTITADESTSTVYMELSSLRSEDTAVYYCARG
LYYDPARADYWGQGTLVTVSS
J703M1S3-19VH
228
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSWYA
ISWVRQAPGQGLEWMGGITPILGSPTYAQKFQG
RVTITADESTSTVYMELSSLRSEDTAVYYCARG
VYYDPKRADYWGQGTLVTVSS
J703M1S3-1VH
229
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSWYT
ISWVRQAPGQGLEWMGGIMPILGTPVYAQKFQG
RVTITADESTSTVYMELSSLRSEDTAVYYCARG
VYYDFRRANYWGQGTLVTVSS
J703M1S3-20VH
230
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSWYA
ISWVRQAPGQGLEWMGGITPILGAATYAQKFQG
RVTITADESTSTVYMELSSLRSEDTAVYYCARG
IYYDPKRADYWGQGTLVTVSS
J703M1S3-21VH
231
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSWYA
ISWVRQAPGQGLEWMGGITPILGDPIYAQKFQG
RVTITADESTSTVYMELSSLRSEDTAVYYCARG
IYYDPKRADYWGQGTLVTVSS
J703M1S3-22VH
232
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSWYA
ISWVRQAPGQGLEWMGGITPILGNPIYAQKFQG
RVTITADESTSTVYMELSSLRSEDTAVYYCARG
IYYDYKRADYWGQGTLVTVSS
J703M1S3-25VH
233
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSWYA
ISWVRQAPGQGLEWMGGITPILGSANYAQKFQG
RVTITADESTSTVYMELSSLRSEDTAVYYCARG
LFYDFRRADYWGQGTLVTVSS
J703M1S3-28VH
234
EVQLVQSGAEVKKPGSSVKVSCKASGGTFAWYA
ISWVRQAPGQGLEWMGGITPILGNAIYAQKFQG
RVTITADESTSTVYMELSSLRSEDTAVYYCARG
VYYDPKRADYWGQGTLVTVSS
J703M1S3-29VH
235
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSWYA
ISWVRQAPGQGLEWMGGITPILGNPIYAQKFQG
RVTITADESTSTVYMELSSLRSEDTAVYYCARG
VYYDPKRADYWGQGTLVTVSS
J703M1S3-2VH
236
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSWYA
TSWVRQAPGQGLEWMGGITPILGSPIYAQKFQG
RVTITADESTSTVYMELSSLRSEDTAVYYCARG
VYYDHRRADYWGQGTLVTVSS
J703M1S3-34VH
237
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSWYA
ISWVRQAPGQGLEWMGGITPILGSPIYAQKFQG
RVTITADESTSTVYMELSSLRSEDTAVYYCARG
VYYDYKRADYWGQGTLVTVSS
J703M1S3-37VH
238
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSWYA
ISWVRQAPGQGLEWMGGITPILGSAIYAQKFQG
RVTITADESTSTVYMELSSLRSEDTAVYYCARG
VYYDPKRADYWGQGTLVTVSS
J703M1S3-38VH
239
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSWYA
ISWVRQAPGQGLEWMGGITPILGTPIYAQKFQG
RVTITADESTSTVYMELSSLRSEDTAVYYCARG
VYYDFKRADYWGQGTLVTVSS
J703M1S3-3VH
240
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSWYT
ISWVRQAPGQGLEWMGGIMPILGTPIYAQKFQG
RVTITADESTSTVYMELSSLRSEDTAVYYCARG
VYYDPRRADYWGQGTLVTVSS
J703M1S3-41VH
241
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSWYA
ISWVRQAPGQGLEWMGGITPILGSANYAQKFQG
RVTITADESTSTVYMELSSLRSEDTAVYYCARG
VYYDPKRADYWGQGTLVTVSS
J703M1S3-42VH
242
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSWYA
ISWVRQAPGQGLEWMGGITPILGAPVYAQKFQG
RVTITADESTSTVYMELSSLRSEDTAVYYCARG
VYYDPKRADYWGQGTLVTVSS
J703M1S3-45VH
243
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSWYA
ISWVRQAPGQGLEWMGGITPILGSAIYAQKFQG
RVTITADESTSTVYMELSSLRSEDTAVYYCARG
VYYDPKRADYWGQGTLVTVSS
J703M1S3-46VH
244
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSWYA
ISWVRQAPGQGLEWMGGITPILGSPIYAQKFQG
RVTITADESTSTVYMELSSLRSEDTAVYYCARG
IYYDPKRADYWGQGTLVTVSS
J703M1S3-47VH
245
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSWYT
ISWVRQAPGQGLEWMGGIMPILGSANYAQKFQG
RVTITADESTSTVYMELSSLRSEDTAVYYCARG
IYYDPKRADYWGQGTLVTVSS
J703M1S3-4VH
246
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSWYA
ISWVRQAPGQGLEWMGGITPILGNAIYAQKFQG
RVTITADESTSTVYMELSSLRSEDTAVYYCARG
IYYDPKRADYWGQGTLVTVSS
J703M1S3-50VH
247
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSWYA
ISWVRQAPGQGLEWMGGITPILGAATYAQKFQG
RVTITADESTSTVYMELSSLRSEDTAVYYCARG
VYYDPKRADYWGQGTLVTVSS
J703M1S3-51VH
248
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSWYA
ISWVRQAPGQGLEWMGGITPILGSPIYAQKFQG
RVTITADESTSTVYMELSSLRSEDTAVYYCARG
IYYDYRRADYWGQGTLVTVSS
J703M1S3-53VH
249
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSWYA
ISWVRQAPGQGLEWMGGIMPILGIPTYAQKFQG
RVTITADESTSTVYMELSSLRSEDTAVYYCARG
VYYDPARADYWGQGTLVTVSS
J703M1S3-54VH
250
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSWYA
ISWVRQAPGQGLEWMGGITPILGSPIYAQKFQG
RVTITADESTSTVYMELSSLRSEDTAVYYCARG
VYYDPKRADYWGQGTLVTVSS
J703M1S3-57VH
251
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSWYA
ISWVRQAPGQGLEWMGGITPILGSAVYAQKFQG
RVTITADESTSTVYMELSSLRSEDTAVYYCARG
VYYDPKRADYWGQGTLVTVSS
J703M1S3-5VH
252
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSWYA
ISWVRQAPGQGLEWMGGITPILGSAIYAQKFQG
RVTITADESTSTVYMELSSLRSEDTAVYYCARG
IYYDYKRADYWGQGTLVTVSS
J703M1S3-62VH
253
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSWYA
ISWVRQAPGQGLEWMGGITPILGYPIYAQKFQG
RVTITADESTSTVYMELSSLRSEDTAVYYCARG
VYYDPKRADYWGQGTLVTVSS
J703M1S3-6VH
254
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSWYA
ISWVRQAPGQGLEWMGGITPILGAATYAQKFQG
RVTITADESTSTVYMELSSLRSEDTAVYYCARG
IYYDFRRADYWGQGTLVTVSS
J703M1S3-72VH
255
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSWYP
ISWVRQAPGQGLEWMGGITPILGSAIYAQKFQG
RVTITADESTSTVYMELSSLRSEDTAVYYCARG
VYYDFRRADYWGQGTLVTVSS
J703M1S3-78VH
256
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSWYA
ISWVRQAPGQGLEWMGGITPILGTANYAQKFQG
RVTITADESTSTVYMELSSLRSEDTAVYYCARG
VYYDPKRADYWGQGTLVTVSS
J703M1S3-79VH
257
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSWYA
ISWVRQAPGQGLEWMGGITPILGSAVYAQKFQG
RVTITADESTSTVYMELSSLRSEDTAVYYCARG
IYYDPKRADYWGQGTLVTVSS
J703M1S3-7VH
258
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSWYA
ISWVRQAPGQGLEWMGGITPILGNPIYAQKFQG
RVTITADESTSTVYMELSSLRSEDTAVYYCARG
VYYDPARADYWGQGTLVTVSS
J703M1S3-81VH
259
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSWYT
ISWVRQAPGQGLEWMGGIMPILGAPNYAQKFQG
RVTITADESTSTVYMELSSLRSEDTAVYYCARG
VYYDYTRADYWGQGTLVTVSS
J703M1S3-83VH
260
EVQLVQSGAEVKKPGSSVKVSCKASGGTFAWYA
ISWVRQAPGQGLEWMGGITPILGSPTYAQKFQG
RVTITADESTSTVYMELSSLRSEDTAVYYCARG
VYYDPKRADYWGQGTLVTVSS
J703M1S3-86VH
261
EVQLVQSGAEVKKPGSSVKVSCKASGGTFGWYA
TSWVRQAPGQGLEWMGGIIPILGTPNYAQKFQG
RVTITADESTSTVYMELSSLRSEDTAVYYCARG
VYYDPKRADYWGQGTLVTVSS
J703M1S3-87VH
262
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSWYT
ISWVRQAPGQGLEWMGGIMPILGTPTYAQKFQG
RVTITADESTSTVYMELSSLRSEDTAVYYCARG
VYYDPKRADYWGQGTLVTVSS
J703M1S3-88VH
263
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSWYT
ISWVRQAPGQGLEWMGGIMPILGSPNYAQKFQG
RVTITADESTSTVYMELSSLRSEDTAVYYCARG
IYYDPKRADYWGQGTLVTVSS
J703M1S3-91VH
264
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSWYA
ISWVRQAPGQGLEWMGGIMPILGSATYAQKFQG
RVTITADESTSTVYMELSSLRSEDTAVYYCARG
VYFDPKRADYWGQGTLVTVSS
J703M1S3-93VH
265
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSWYA
ISWVRQAPGQGLEWMGGITPILGAANYAQKFQG
RVTITADESTSTVYMELSSLRSEDTAVYYCARG
LYYDPKRADYWGQGTLVTVSS
J703M1S3-9VH
266
EVQLVQSGAEVKKPGSSVKVSCKASGGTFSWYP
ISWVRQAPGQGLEWMGGITPILGAGIYAQKFQG
RVTITADESTSTVYMELSSLRSEDTAVYYCARG
VYYDFKRADYWGQGTLVTVSS
Table 12 provides a list of amino acid sequences of VL regions of affinity matured fully human TNF antibodies derived from AE11-5. Amino acid residues of individual CDRs of each VH sequence are indicated in bold.
TABLE 12
List of amino acid sequences of affinity matured
AE11-5 VL variants
Clone
SEQ ID NO:
VL
J685M2S2-17Vk
267
EIVLTQSPDFQSVTPKEKVTITCRASQSIGSSLH
WYQQKPDQSPKLLIKHASQSISGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQSSSSPPPTFGQG
TKVEIK
J685M2S2-94Vk
268
EIVLTQSPDFQSVTPKEKVTITCRASQSIGSSLH
WYQQKPDQSPKLLIKHASQSISGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQSSSSPPPTFGQW
TKVEIK
J688M2-37Vk
269
EIVLTQSPDFQSVTPKEKVTITCRARQSIGSSLH
WYQQKPDQSPKLLIKHASQSISGVPSRFSGSGSG
TNFTLTINSLEAEDAATYYCHQSSSSPPPTFGQG
TKVEIK
J688M2-90Vk
270
EIVLTQSPDFQSVTPKEKVTITCRASQSIGSSLH
WYQQKPDQSPKLLIKHASQSISGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQSSSSPPPTFGQG
TKVEIK
J693FRM2S2L-
271
EIVLTQSPDFQSVTPKEKVTITCRASQSIGRSLH
26Vk
WYQQKPDQSPKLLIKHASQSVSGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQNRSSPPSTFGQG
TKVEIK
J693FRM2S2L-
272
EIVLTQSPDFQSVTPKEKVTITCRASQSIGRSLH
27Vk
WYQQKPDQSPKLLIKYASQSLSGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQSSSSPPVTFGQG
TKVEIK
J693FRM2S2L-
273
EIVLTQSPDFQSVTPKEKVTITCRASQSIGSSLH
29Vk
WYQQKPDQSPKLLIKYASQSISGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQRSNLPAPTFGQG
TKVEIK
J693FRM2S2L-
274
EIVLTQSPDFQSVTPKEKVTITCRASQIIGGSLH
39Vk
WYQQKPDQSPKLLIKYASQSFSGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQPICSPPRTFGQG
TKVEIK
J693FRM2S2L-
275
EIVLTQSPDFQSVTPKEKVTITCRASQTIGSNLH
3Vk
WYQQKPDQSPKLLIKYASQSLSGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQCSISPPATFGQG
TKVEIK
J693FRM2S2L-
276
EIVLTQSPDFQSVTPKEKVTITCRASQCIGTSLH
40Vk
WYQQKPDQSPKLLIKYDSQSISGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQNSSSPPPTFGQG
TKVEIK
J693FRM2S2L-
277
EIVLTQSPDFQSVTPKEKVTITCRASQNIGNSLH
42Vk
WYQQKPDQSPKLLIKYTSQSISGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQTSSLPLPTFGQG
TKVEIK
J693FRM2S2L-
278
EIVLTQSPDFQSVTPKEKVTITCRASQSIGSSLH
43Vk
WYQQKPDQSPKLLIKYVSQSISGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQISDLPTSTFGQG
TKVEIK
J693FRM2S2L-
279
EIVLTQSPDFQSVTPKEKVTITCRASQRIGSNLH
45Vk
WYQQKPDQSPKLLIKYASQSISGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQGSSLPPPTFGQG
TKVEIK
J693FRM2S2L-
280
EIVLTQSPDFQSVTPKEKVTITCRASQCIGSSLH
46Vk
WYQQKPDQSPKLLIKHTSQSNSGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQSNSSPLSTFGQG
TKVEIK
J693FRM2S2L-
281
EIVLTQSPDFQSVTPKEKVTITCRASQNIGGSLH
47Vk
WYQQKPDQSPKLLIKYASQSISGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQNSSLPLPTFGQG
TKVEIK
J693FRM2S2L-
282
EIVLTQSPDFQSVTPKEKVTITCRASQSIGSRLH
48Vk
WYQQKPDQSPKLLIKYASQSISGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQSSKSPPPTFGQG
TKVEIK
J693FRM2S2L-
283
EIVLTQSPDFQSVTPKEKVTITCRASQSIGSCLH
52Vk
WYQQKPDQSPKLLIKYASQSVSGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQRSSLPTPTFGQG
TKVEIK
J693FRM2S2L-
284
EIVLTQSPDFQSVTPKEKVTITCRASQSIGGRLH
53Vk
WYQQKPDQSPKLLIKYASQSLSGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQASSSPSTTFGQG
TKVEIK
J693FRM2S2L-
285
EIVLTQSPDFQSVTPKEKVTITCRASQRIGPSLH
54Vk
WYQQKPDQSPKLLIKYASQSISGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQNSCLPSTTFGQG
TKVEIK
J693FRM2S2L-
286
EIVLTQSPDFQSVTPKEKVTITCRASQSIGRSLH
58Vk
WYQQKPDQSPKLLIKYASQSRSGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQSGISPPTTFGQG
TKVEIK
J693FRM2S2L-
287
EIVLTQSPDFQSVTPKEKVTITCRASQSIGTSLH
59Vk
WYQQKPDQSPKLLIKYVSQSISGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQGMSSPAPTFGQG
TKVEIK
J693FRM2S2L-
288
EIVLTQSPDFQSVTPKEKVTITCRASQSIGSSLH
5Vk
WYQQKPDQSPKLLIKYASQSLSGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQRRNSPPPTFGQG
TKVEIK
J693FRM2S2L-
289
EIVLTQSPDFQSVTPKEKVTITCRASQKIGSGLH
88Vk
WYQQKPDQSPKLLIKHASQSISGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQNNSSPHKTFGQG
TKVEIK
J693FRM2S2L-
290
EIVLTQSPDFQSVTPKEKVTITCRASQTIGSNLH
89Vk
WYQQKPDQSPKLLIKHSSQSISGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQNSSSPLPTFGQG
TKVEIK
J693FRM2S2L-
291
EIVLTQSPDFQSVTPKEKVTITCRASQNIGRSLH
8Vk
WYQQKPDQSPKLLIKYASQSSSGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQSRSSPPPTFGQG
TKVEIK
J693FRM2S2L-
292
EIVLTQSPDFQSVTPKEKVTITCRASQCIGKSLH
90Vk
WYQQKPDQSPKLLIKHPSQSVSGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQSIGLPPTTFGQG
TKVEIK
J693FRM2S2L-
293
EIVLTQSPDFQSVTPKEKVTITCRASQTIGSSLH
91Vk
WYQQKPDQSPKLLIKHASQSLSGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQSSISPPATFGQG
TKVEIK
J693FRM2S2L-
294
EIVLTQSPDFQSVTPKEKVTITCRASQSIGSTLH
92Vk
WYQQKPDQSPKLLIKYESQSISGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQRCCSPTQTFGQG
TKVEIK
J693FRM2S2L-
295
EIVLTQSPDFQSVTPKEKVTITCRASQSIGRKLH
94Vk
WYQQKPDQSPKLLIKYSSQSISGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQSSRSPPTTFGQG
TKVEIK
J693FRM2S2R-
296
EIVLTQSPDFQSVTPKEKVTITCRASQTIGTSLH
10Vk
WYQQKPDQSPKLLIKHASQSLSGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQSSSSPSPTFGQG
TKVEIK
J693FRM2S2R-
297
EIVLTQSPDFQSVTPKEKVTITCRASQSIGSSLH
11Vk
WYQQKPDQSPKLLIKHVSQSISGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQRGSSPPRTFGQG
TKVEIK
J693FRM2S2R-
298
EIVLTQSPDFQSVTPKEKVTITCRASQTIGSTLH
12Vk
WYQQKPDQSPKLLIKHTSQSISGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQRSSSPPPTFGQG
TKVEIK
J693FRM2S2R-
299
EIVLTQSPDFQSVTPKEKVTITCRASQSIGSNLH
14Vk
WYQQKPDQSPKLLIKHGSQSISGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQRHSSPRATFGQG
TKVEIK
J693FRM2S2R-
300
EIVLTQSPDFQSVTPKEKVTITCRASQKIGSNLH
15Vk
WYQQKPDQSPKLLIKYASQSFSGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQNSSSPPATFGQG
TKVEIK
J693FRM2S2R-
301
EIVLTQSPDFQSVTPKEKVTITCRASQSIGRSLH
16Vk
WYQQKPDQSPKLLIKYASQSISGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQSSRSPFRTFGQG
TKVEIK
J693FRM2S2R-
302
EIVLTQSPDFQSVTPKEKVTITCRASQCIGRRLH
34Vk
WYQQKPDQSPKLLIKHASQSRSGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQCTSSPPPTFGQG
TKVEIK
J693FRM2S2R-
303
EIVLTQSPDFQSVTPKEKVTITCRASQRIGSNLH
36Vk
WYQQKPDQSPKLLIKHASQSISGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQSLRLPPQTFGQG
TKVEIK
J693FRM2S2R-
304
EIVLTQSPDFQSVTPKEKVTITCRASQSIGSSLH
39Vk
WYQQKPDQSPKLLIKHASQSISGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQNRSLPRLTFGQG
TKVEIK
J693FRM2S2R-
305
EIVLTQSPDFQSVTPKEKVTITCRASQSIGSCLH
3Vk
WYQQKPDQSPKLLIKYASQSISGVPSSSVASGSG
TDFTLTINSLEAEDAATYYCHQRSSLPQPTFGQG
TKVEIK
J693FRM2S2R-
306
EIVLTQSPDFQSVTPKEKVTITCRASQSIGRRLH
42Vk
WYQQKPDQSPKLLIKHPSQSVSGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQSIDSPPPTFGQG
TKVEIK
J693FRM2S2R-
307
EIVLTQSPDFQSVTPKEKVTITCRASQTIGRSLH
45Vk
WYQQKPDQSPKLLIKYKSQSSSGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQRWGLPMPTFGQG
TKVEIK
J693FRM2S2R-
308
EIVLTQSPDFQSVTPKEKVTITCRASQRIGSMLH
48Vk
WYQQKPDQSPKLLIKHSSQSISGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQTNSLPPRTFGQG
TKVEIK
J693FRM2S2R-
309
EIVLTQSPDFQSVTPKEKVTITCRASQSIGRSLH
50Vk
WYQQKPDQSPKLLIKHASQSISGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQGSRSPLDTFGQG
TKVEIK
J693FRM2S2R-
310
EIVLTQSPDFQSVTPKEKVTITCRASQSIGCSLH
51Vk
WYQQKPDQSPKLLIKYASQSVSVVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQSSTLPPPTFGQG
TKVEIK
J693FRM2S2R-
311
EIVLTQSPDFQSVTPKEKVTITCRASQGIGTSLH
52Vk
WYQQKPDQSPKLLIKHDSQSISGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQTSSLPPPTFGQG
TKVEIK
J693FRM2S2R-
312
EIVLTQSPDFQSVTPKEKVTITCRASQIIGSSLH
56Vk
WYQQKPDQSPKLLIKHASQSISGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQSSSLPLPTFGQG
TKVEIK
J693FRM2S2R-
313
EIVLTQSPDFQSVTPKEKVTITCRASQSIGSSLH
58Vk
WYQQKPDQSPKLLIKYTSQSKSGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQGNRSPSTTFGQG
TKVEIK
J693FRM2S2R-
314
EIVLTQSPDFQSVTPKEKVTITCRASKRIGSSLH
59Vk
WYQQKPDQSPKLLIKHKSQSISGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQRSASPPPTFGQG
TKVEIK
J693FRM2S2R-
315
EIVLTQSPDFQSVTPKEKVTITCRASQTIGSSLH
5Vk
WYQQKPDQSPKLLIKHPSQSMSGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQSSTSPPATFGQG
TKVEIK
J693FRM2S2R-
316
EIVLTQSPDFQSVTPKEKVTITCRASQSIGSSLH
60Vk
WYQQKPDQSPKLLIKHASQSISGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQRSSLPTPTFGQG
TKVEIK
J693FRM2S2R-
317
EIVLTQSPDFQSVTPKEKVTITCRASQSIGSTLH
61Vk
WYQQKPDQSPKLLIKHASQSFSGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQSNCSPAHTFGQG
TKVEIK
J693FRM2S2R-
318
EIVLTQSPDFQSVTPKEKVTITCRASQTIGSRLH
62Vk
WYQQKPDQSPKLLIKYVSQSISGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQGSRLPPPTFGQG
TKVEIK
J693FRM2S2R-
319
EIVLTQSPDFQSVTPKEKVTITCRASQRIGSTLH
63Vk
WYQQKPDQSPKLLIKHASQSNSGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQSSCSPQATFGQG
TKVEIK
J693FRM2S2R-
320
EIVLTQSPDFQSVTPKEKVTITCRASQSIGTSLH
64Vk
WYQQKPDQSPKLLIKYPSQSISGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQSGRSPPHTFGQG
TKVEIK
J693FRM2S2R-
321
EIVLTQSPDFQSVTPKEKVTITCRASQSIGSSLH
65Vk
WYQQKPDQSPKLLIKHASQSISGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQNSILPPPTFGQG
TKVEIK
J693FRM2S2R-
322
EIVLTQSPDFQSVTPKEKVTITCRASQCIGSYLH
92Vk
WYQQKPDQSPKLLIKHVSQSISGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQSSSSPTLTFGQG
TKVEIK
J693FRM2S2R-
323
EIVLTQSPDFQSVTPKEKVTITCRASQSIGSSLH
93Vk
WYQQKPDQSPKLLIKHASQSMSGVPSGFSGSGSG
TDFTLTINSLEAEDAATYYCHQTNRSPPPTFGQG
TKVEIK
J693FRM2S2R-
324
EIVLTQSPDFQSVTPKEKVTITCRASQNIGTSLH
9Vk
WYQQKPDQSPKLLIKYVSQSISGVPSRFSGSGSG
TDFTLNINSLEAEDAATYYCHQSSCLPRPTFGQG
TKVEIK
J693M2S2L-10Vk
325
EIVLTQSPDFQSVTPKEKVTITCRASQSIGSPLH
WYQQKPDQSPKLLIKYASQSISGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQGSSSPPPTFGQG
TKVEIK
J693M2S2L-11Vk
326
EIVLTQSPDFQSVTPKEKVTITCRASQSIGSTLH
WYQQKPDQSPKLLIKHDSQSKSGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQSSDSPAPTFGQG
TKVEIK
J693M2S2L-12Vk
327
EIVLTQSPDFQSVTPKEKVTITCRASQSIGSCLH
WYQQKPDQSPKLLIKHASQSNSGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQSRISPLPTFGQG
TKVEIK
J693M2S2L-13Vk
328
EIVLTQSPDFQSVTPKEKVTITCRASQSIGRRLH
WYQQKPDQSPKLLIKHSSQSISGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQCSSLPHPTFGQG
TKVEIK
J693M2S2L-14Vk
329
EIVLTQSPDFQSVTPKEKVTITCRASQRIGSRLH
WYQQKPDQSPKLLIKHASQSTSGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQSCSSPLVTFGQG
TKVEIK
J693M2S2L-16Vk
330
EIVLTQSPDFQSVTPKEKVTITCRASQSIGSRLH
WYQQKPDQSPKLLIKHASQSSSGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQSGSSPQATFGQG
TKVEIK
J693M2S2L-17Vk
331
EIVLTQSPDFQSVTPKEKVTITCRASQSIGSSLH
WYQQKPDQSPKLLIKHASQSVSGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQNRGSPPQTFGQG
TKVEIK
J693M2S2L-18Vk
332
EIVLTQSPDFQSVTPKEKVTITCRASQTIGSILH
WYQQKPDQSPKLLIKHASQSISGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQNTSLPPPTFGQG
TKVEIK
J693M2S2L-19Vk
333
EIVLTQSPDFQSVTPKEKVTITCRASQSIGNSLH
WYQQKPDQSPKLLIKYPSQSISGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQGSRLPVPTFGQG
TKVEIK
J693M2S2L-1Vk
334
EIVLTQSPDFQSVTPKEKVTITCRASQSIGSRLH
WYQQKPDQSPKLLIKHTSQSNSGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQSSSLPAPTFGQG
TKVEIK
J693M2S2L-20Vk
335
EIVLTQSPDFQSVTPKEKVTITCRASQNIGSSLH
WYQQKPDQSPKLLIKHVSQSISGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQSNSLPAPTFGQG
TKVEIK
J693M2S2L-21Vk
336
EIVLTQSPDFQSVTPKEKVTITCRASQSIGSSLH
WYQQKPDQSPKLLIKHASQSISGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQSMSLPSATFGQG
TKVEIK
J693M2S2L-22Vk
337
EIVLTQSPDFQSVTPKEKVTITCRASQRIGSSLH
WYQQKPDQSPKLLIKHLSQSISGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQPCRLPPSTFGQG
TKVEIK
J693M2S2L-23Vk
338
EIVLTQSPDFQSVTPKEKVTITCRASQSIGSLLH
WYQQKPDQSPKLLIKHASQSISGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQSCSSPRHTFGQG
TKVEIK
J693M2S2L-24Vk
339
EIVLTQSPDFQSVTPKEKVTITCRASQRIGSSLH
WYQQKPDQSPKLLIKHPSQSKSGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQSSRSPAPTFGQG
TKVEIK
J693M2S2L-25Vk
340
EIVLTQSPDFQSVTPKEKVTITCRASQSIGGSLH
WYQQKPDQSPKLLIKYSSQSLSGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQSSILPSLTFGQG
TKVEIK
J693M2S2L-26Vk
341
EIVLTQSPDFQSVTPKEKVTITCRASQSIGSSLH
WYQQKPDQSPKLLIKHPSQSISGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQSRNLPPRTFGQG
TKVEIK
J693M2S2L-27Vk
342
EIVLTQSPDFQSVTPKEKVTITCRASQSIGSILH
WYQQKPDQSPKLLIKYGSQSLSGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQNGSSPPRTFGQG
TKVEIK
J693M2S2L-28Vk
343
EIVLTQSPDFQSVTPKEKVTITCRASQSIGSSLH
WYQQKPDQSPKLLIKYFSQSISGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQNSCLPMQTFGQG
TKVEIK
J693M2S2L-29Vk
344
EIVLTQSPDFQSVTPKEKVTITCRASQNIGSSLH
WYQQKPDQSPKLLIKYSSQSVSGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQSSISPPATFGQG
TKVEIK
J693M2S2L-2Vk
345
EIVLTQSPDFQSVTPKEKVTITCRASQCIGSSLH
WYQQKPDQSPKLLIKHASQSNSGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQSTCLPPRTFGQG
TKVEIK
J693M2S2L-30Vk
346
EIVLTQSPDFQSVTPKEKVTITCRASQSIGSRLH
WYQQKPDQSPKLLIKYVSQSMSGVLSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQPSTSPRPTFGQG
TKVEIK
J693M2S2L-31Vk
347
EIVLTQSPDFQSVTPKEKVTITCRASQSIGSRLH
WYQQKPDQSPKLLIKHASQSISGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQNSSLPPSTFGQG
TKVEIK
J693M2S2L-32Vk
348
EIVLTQSPDFQSVTPKEKVTITCRASQSIGCSLH
WYQQKPDQSPKLLIKYASQSNSGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQSSSSPSSTFGQG
TKVEIK
J693M2S2L-33Vk
349
EIVLTQSPDFQSVTPKEKVTITCRASQIIGTSLH
WYQQKPDQSPKLLIKYASQSISGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQSSRSPPRTFGQG
TKVEIK
J693M2S2L-34Vk
350
EIVLTQSPDFQSVTPKEKVTITCRASQKIGTSLH
WYQQKPDQSPKLLIKHESQSISGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQSSGSPPPTFGQG
TKVEIK
J693M2S2L-35Vk
351
EIVLTQSPDFQSVTPKEKVTITCRASQTIGGSLH
WYQQKPDQSPKLLIKHVSQSVSGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQSSISPPPTFGQG
TKVEIK
J693M2S2L-36Vk
352
EIVLTQSPDFQSVTPKEKVTITCRASQSIGSDLH
WYQQKPDQSPKLLIKHVSQSVSGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQSCMSPSLTFGQG
TKVEIK
J693M2S2L-37Vk
353
EIVLTQSPDFQSVTPKEKVTITCRASQSIGSNLH
WYQQKPDQSPKLLIKHASQSISGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQSSSLPNTTFGQG
TKVEIK
J693M2S2L-38Vk
354
EIVLTQSPDFQSVTPKEKVTITCRASQRIGSILH
WYQQKPDQSPKLLIKHASQSISGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQGRISPSSTFGQG
TKVEIK
J693M2S2L-39Vk
355
EIVLTQSPDFQSVTPKEKVTITCRASQSIGNRLH
WYQQKPDQSPKLLIKHASQSLSGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQSGSLPTLTFGQG
TKVEIK
J693M2S2L-3Vk
356
EIVLTQSPDFQSVTPKEKVTITCRASQTIGSSLH
WYQQKPDQSPKLLIKHDSQSLSGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQNSSLPTHTFGQG
TKVEIK
J693M2S2L-40Vk
357
EIVLTQSPDFQSVTPKEKVTITCRASQTIGRSLH
WYQQKPDQSPKLLIKHGSQSISGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQSRSSPPSTFGQG
TKVEIK
J693M2S2L-41Vk
358
EIVLTQSPDFQSVTPKEKVTITCRASQSIGSSLH
WYQQKPDQSPKLLIKYASQSISGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQNCSSPPPTFGQG
TKVEIK
J693M2S2L-44Vk
359
EIVLTQSPDFQSVTPKEKVTITCRASQSIGSSLH
WYQQKPDQSPKLLIKYESQSDSGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQRRNSPPSTFGQG
TKVEIK
J693M2S2L-45Vk
360
EIVLTQSPDFQSVTPKEKVTITCRASQGIGSRLH
WYQQKPDQSPKLLIKHGSQSISGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQNRGLPAPTFGQG
TKVEIK
J693M2S2L-46Vk
361
EIVLTQSPDFQSVTPKEKVTITCRASQSIGSSLH
WYQQKPDQSPKLLIKYASQSSSGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQNHTSPPPTFGQG
TKVEIK
J693M2S2L-47Vk
362
EIVLTQSPDFQSVTPKEKVTITCRASQSIGSRLH
WYQQKPDQSPKLLIKHASQSVSGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQSGRLPPPTFGQG
TKVEIK
J693M2S2L-4Vk
363
EIVLTQSPDFQSVTPKEKVTITCRASQYIGKRLH
WYQQKPDQSPKLLIKYASQSISGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQSNISPPPTFGQG
TKVEIK
J693M2S2L-51Vk
364
EIVLTQSPDFQSVTPKEKVTITCRASQRIGSSLH
WYQQKPDQSPKLLIKHESQSISGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQSSSLPPPTFGQG
TKVEIK
J693M2S2L-52Vk
365
EIVLTQSPDFQSVTPKEKVTITCRASQTIGSSLH
WYQQKPDQSPKLLIKYASQSISGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQRSSLPPSTFGQG
TKVEIK
J693M2S2L-54Vk
366
EIVLTQSPDFQSVTPKEKVTITCRASQSIGRSLH
WYQQKPDQSPKLLIKHPSQSISGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQCSSSPAQTFGQG
TKVEIK
J693M2S2L-55Vk
367
EIVLTQSPDFQSVTPKEKVTITCRASQSIGRSLH
WYQQKPDQSPKLLIKHTSQSLSGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQRSSLPLPTFGQG
TKVEIK
J693M2S2L-56Vk
368
EIVLTQSPDFQSVTPKEKVTITCRASQWIGSSLH
WYQQKPDQSPKLLIKHTSQSISGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQSSSLPPQTFGQG
TKVEIK
J693M2S2L-58Vk
369
EIVLTQSPDFQSVTPKEKVTITCRASQSIGRSLH
WYQQKPDQSPKLLIKYSSQSISGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQGSSSPPPTFGQG
TKVEIK
J693M2S2L-59Vk
370
EIVLTQSPDFQSVTPKEKVTITCRASQSIGSSLH
WYQQKPDQSPKLLIKHASQSISGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQSSRLPPSTFGQG
TKVEIK
J693M2S2L-5Vk
371
EIVLTQSPDFQSVTPKEKVTITCRASQSIGSSLH
WYQQKPDQSPKLLIKYGSQSISGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQNWSLPLPTFGQG
TKVEIK
J693M2S2L-62Vk
372
EIVLTQSPDFQSVTPKEKVTITCRASQRIGTSLH
WYQQKPDQSPKLLIKYASQSKSGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQSSCSPTPTFGQG
TKVEIK
J693M2S2L-64Vk
373
EIVLTQSPDFQSVTPKEKVTITCRASQSIGGSLH
WYQQKPDQSPKLLIKYGSQSISGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQRCVSPSPTFGQG
TKVEIK
J693M2S2L-65Vk
374
EIVLTQSPDFQSVTPKEKVTITCRASQSIGGTLH
WYQQKPDQSPKLLIKYASQSISGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQSSSSPARTFGQG
TKVEIK
J693M2S2L-66Vk
375
EIVLTQSPDFQSVTPKEKVTITCRASQSIGSSLH
WYQQKPDQSPKLLIKYASQSISGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQCGSSPLHTFGQG
TKVEIK
J693M2S2L-67Vk
376
EIVLTQSPDFQSVTPKEKVTITCRASQSIGTSLH
WYQQKPDQSPKLLIKHPSQSLSGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQSTSSPPPTFGQG
TKVEIK
J693M2S2L-68Vk
377
EIVLTQSPDFQSVTPKEKVTITCRASQNIGSSLH
WYQQKPDQSPKLLIKHASQSISGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQNSGLPLPTFGQG
TKVEIK
J693M2S2L-69Vk
378
EIVLTQSPDFQSVTPKEKVTITCRASQSIGRRLH
WYQQKPDQSPKLLIKYASQSISGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQGSSSPSPTFGQG
TKVEIK
J693M2S2L-6Vk
379
EIVLTQSPDFQSVTPKEKVTITCRASQRIGGNLH
WYQQKPDQSPKLLIKHESQSNSGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQSSSLPSHTFGQG
TKVEIK
J693M2S2L-70Vk
380
EIVLTQSPDFQSVTPKEKVTITCRASQSIGSSLH
WYQQKPDQSPKLLIKYASQSTSGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQCSSSPSHTFGQG
TKVEIK
J693M2S2L-71Vk
381
EIVLTQSPDFQSVTPKEKVTITCRASQSIGSSLH
WYQQKPDQSPKLLIKHASQSMSGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQSRNSPPTTFGQG
TKVEIK
J693M2S2L-72Vk
382
EIVLTQSPDFQSVTPKEKVTITCRASQRIGSRLH
WYQQKPDQSPKLLIKHGSQSISGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQNSSSPPPTFGQG
TKVEIK
J693M2S2L-74Vk
383
EIVLTQSPDFQSVTPKEKVTITCRASQNIGSSLH
WYQQKPDQSPKLLIKYASQSLSGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQSSLLPAPTFGQG
TKVEIK
J693M2S2L-75Vk
384
EIVLTQSPDFQSVTPKEKVTITCRASQIIGTTLH
WYQQKPDQSPKLLIKHASQSISGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQNSNLPPSTFGQG
TKVEIK
J693M2S2L-76Vk
385
EIVLTQSPDFQSVTPKEKVTITCRASQNIGGNLH
WYQQKPDQSPKLLIKHASQSLSGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQSSNLPPPTFGQG
TKVEIK
J693M2S2L-77Vk
386
EIVLTQSPDFQSVTPKEKVTITCRASQGIGGSLH
WYQQKPDQSPKLLIKYASQSTSGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQSACLPTRTFGQG
TKVEIK
J693M2S2L-78Vk
387
EIVLTQSPDFQSVTPKEKVTITCRASQSIGTSLH
WYQQKPDQSPKLLIKYASQSVSGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQIGSLPPPTFGQG
TKVEIK
J693M2S2R-13Vk
388
EIVLTQSPDFQSVTPKEKVTITCRASQSIGRSLH
WYQQKPDQSPKLLIKHASQSVSGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQNSRLPPPTFGQG
TKVEIK
J693M2S2R-14Vk
389
EIVLTQSPDFQSVTPKEKVTITCRASQSIGSSLH
WYQQKPDQSPKLLIKHNSQSISGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQRSSSPPLTFGQG
TKVEIK
J693M2S2R-15Vk
390
EIVLTQSPDFQSVTPKEKVTITCRASQSIGRNLH
WYQQKPDQSPKLLIKHVSQSISGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQRSRSPPSTFGQG
TKVEIK
J693M2S2R-16Vk
391
EIVLTQSPDFQSVTPKEKVTITCRASQSIGSSLH
WYQQKPDQSPKLLIKHASQSVSGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQCSSLPAPTFGQG
TKVEIK
J693M2S2R-17Vk
392
EIVLTQSPDFQSVTPKEKVTITCRASQRIGSSLH
WYQQKPDQSPKLLIKHASQSLSGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQSSRLPPQTFGQG
TKVEIK
J693M2S2R-18Vk
393
EIVLTQSPDFQSVTPKEKVTITCRASQCIGSRLH
WYQQKPDQSPKLLIKYASQSISGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQRGRLPPRTFGQG
TKVEIK
J693M2S2R-19Vk
394
EIVLTQSPDFQSVTPKEKVTITCRASQSIGSSLH
WYQQKPDQSPKLLIKYASQSISGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQSTSLPRLTFGQG
TKVEIK
J693M2S2R-20Vk
395
EIVLTQSPDFQSVTPKEKVTITCRASQIIGSSLH
WYQQKPDQSPKLLIKYASQSISGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQSRSSPQQTFGQG
TKVEIK
J693M2S2R-21Vk
396
EIVLTQSPDFQSVTPKEKVTITCRASQSIGSTLH
WYQQKPDQSPKLLIKHASQSISGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQSSSLPPPTFGQG
TKVEIK
J693M2S2R-22Vk
397
EIVLTQSPDFQSVTPKEKVTITCRASQSIGNSLH
WYQQKPDQSPKLLIKHGSQSISGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQRRSSPRHTFGQG
TKVEIK
J693M2S2R-27Vk
398
EIVLTQSPDFQSVTPKEKVTITCRASQRIGRRLH
WYQQKPDQSPKLLIKHASQSISGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQSIGSPPLTFGQG
TKVEIK
J693M2S2R-29Vk
399
EIVLTQSPDFQSVTPKEKVTITCRASQSIGRGLH
WYQQKPDQSPKLLIKYGSQSMSGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQSSSLPPPTFGQG
TKVEIK
J693M2S2R-2Vk
400
EIVLTQSPDFQSVTPKEKVTITCRASQSIGCSLH
WYQQKPDQSPKLLIKYASQSISGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQCTSLPLPTFGQG
TKVEIK
J693M2S2R-30Vk
401
EIVLTQSPDFQSVTPKEKVTITCRASQGIGSSLH
WYQQKPDQSPKLLIKYVSQSISGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQGSSLPTPTFGQG
TKVEIK
J693M2S2R-31Vk
402
EIVLTQSPDFQSVTPKEKVTITCRASQSIGTSLH
WYQQKPDQSPKLLIKHASQSSSGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQSSRLPPLTFGQG
TKVEIK
J693M2S2R-32Vk
403
EIVLTQSPDFQSVTPKEKVTITCRASQVIGGVLH
WYQQKPDQSPKLLIKYTSQSISGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQSSSSPRPTFGQG
TKVEIK
J693M2S2R-33Vk
404
EIVLTQSPDFQSVTPKEKVTITCRASQSIGSSLH
WYQQKPDQSPKLLIKHSSQSLSGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQRSNSPHRTFGQG
TKVEIK
J693M2S2R-36Vk
405
EIVLTQSPDFQSVTPKEKVTITCRASQSIGRTLH
WYQQKPDQSPKLLIKYASQSISGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQCSISPQPTFGQG
TKVEIK
J693M2S2R-37Vk
406
EIVLTQSPDFQSVTPKEKVTITCRASQRIGNTLH
WYQQKPDQSPKLLIKYPSQSISGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQSGSSPPPTFGQG
TKVEIK
J693M2S2R-39Vk
407
EIVLTQSPDFQSVTPKEKVTITCRASQSIGSRLH
WYQQKPDQSPKLLIKYISQSMSGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQSCGLPPPTFGQG
TKVEIK
J693M2S2R-3Vk
408
EIVLTQSPDFQSVTPKEKVTITCRASQNIGTRLH
WYQQKPDQSPKLLIKYGSQSLSGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQSRISPPPTFGQG
TKVEIK
J693M2S2R-40Vk
409
EIVLTQSPDFQSVTPKEKVTITCRASQSIGSTLH
WYQQKPDQSPKLLIKYVSQSISGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQCSRLPPPTFGQG
TKVEIK
J693M2S2R-44Vk
410
EIVLTQSPDFQSVTPKEKVTITCRASQSIGSRLH
WYQQKPDQSPKLLIKYASQSTSGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQSSNLPSPTFGQG
TKVEIK
J693M2S2R-45Vk
411
EIVLTQSPDFQSVTPKEKVTITCRASQSIGSNLH
WYQQKPDQSPKLLIKHASQSMSGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQSSSLPRPTFGQG
TKVEIK
J693M2S2R-46Vk
412
EIVLTQSPDFQSVTPKEKVTITCRASQIIGSSLH
WYQQKPDQSPKLLIKYASQSISGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQSISSPSPTFGQG
TKVEIK
J693M2S2R-47Vk
413
EIVLTQSPDFQSVTPKEKVTITCRASQSIGSSLH
WYQQKPDQSPKLLIKYASQSFSGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQSNCLPPPTFGQG
TKVEIK
J693M2S2R-48Vk
414
EIVLTQSPDFQSVTPKEKVTITCRASQSIGKSLH
WYQQKPDQSPKLLIKHESQSISGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQANSLPPPTFGQG
TKVEIK
J693M2S2R-4Vk
415
EIVLTQSPDFQSVTPKEKVTITCRASQSIGRRLH
WYQQKPDQSPKLLIKYASQSISGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQCSSSPPSTFGQG
TKVEIK
J693M2S2R-52Vk
416
EIVLTQSPDFQSVTPKEKVTITCRASQIIGHSLH
WYQQKPDQSPKLLIKHASQSILGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQSIKSPPATFGQG
TKVEIK
J693M2S2R-54Vk
417
EIVLTQSPDFQSVTPKEKVTITCRASQSIGTSLH
WYQQKPDQSPKLLIKHTSQSKSGVPSRFSGSGSG
TDFALTINSLEAEDAATYYCHQSSNSPRYTFGQG
TKVEIK
J693M2S2R-55Vk
418
EIVLTQSPDFQSVTPKEKVTITCRASQSIGSSLH
WYQQKPDQSPKLLIKHASQSHSGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQSGGSPPWTFGQG
TKVEIK
J693M2S2R-56Vk
419
EIVLTQSPDFQSVTPKEKVTITCRASQGIGRSLH
WYQQKPDQSPKLLIKYASQSLSGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQSNRSPPPTFGQG
TKVEIK
J693M2S2R-5Vk
420
EIVLTQSPDFQSVTPKEKVTITCRASQSIGTTLH
WYQQKPDQSPKLLIKHVSQSTSGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQSSSLPHPTFGQG
TKVEIK
J693M2S2R-60Vk
421
EIVLTQSPDFQSVTPKEKVTITCRASQIIGSSLH
WYQQKPDQSPKLLIKYPSQSTSGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQSWSSPLMTFGQG
TKVEIK
J693M2S2R-61Vk
422
EIVLTQSPDFQSVTPKEKVTITCRASQSIGNTLH
WYQQKPDQSPKLLIKHASQSISGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQSSSSPPPTFGQG
TKVEIK
J693M2S2R-62Vk
423
EIVLTQSPDFQSVTPKEKVTITCRASQRIGICLH
WYQQKPDQSPKLLIKYASQSMSGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQGFSLPPATFGQG
TKVEIK
J693M2S2R-63Vk
424
EIVLTQSPDFQSVTPKEKVTITCRASQSIGSCLH
WYQQKPDQSPKLLIKYPSQSTSGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQGSCSPTTTFGQG
TKVEIK
J693M2S2R-64Vk
425
EIVLTQSPDFQSVTPKEKVTITCRASQRIGNTLH
WYQQKPDQSPKLLIKYPSQSISGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQSGSSPPPTFGQG
TKVEIK
J693M2S2R-65Vk
426
EIVLTQSPDFQSVTPKEKVTITCRASQTIGTSLH
WYQQKPDQSPKLLIKYASQSTSGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQRCSLPPPTFGQG
TKVEIK
J693M2S2R-68Vk
427
EIVLTQSPDFQSVTPKEKVTITCRASQSIGGSLH
WYQQKPDQSPKLLIKYASQSHSGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQCRISPRPTFGQG
TKVEIK
J693M2S2R-69Vk
428
EIVLTQSPDFQSVTPKEKVTITCRASQRIGSSLH
WYQQKPDQSPKLLIKHPSQSKSGVPSRFSGSGSG
TDFTLSINSLEAEDAATYYCHQTSRSPLHTFGQG
TKVEIK
J693M2S2R-6Vk
429
EIVLTQSPDFQSVTPKEKVTITCRASQNIGKNLH
WYQQKPDQSPKLLIKYPSQSISGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQSRSSPLSTFGQG
TKVEIK
J693M2S2R-70Vk
430
EIVLTQSPDFQSVTPKEKVTITCRASQSIGSRLH
WYQQKPDQSPKLLIKYMSQSISGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQSRVLPPPTFGQG
TKVEIK
J693M2S2R-71Vk
431
EIVLTQSPDFQSVTPKEKVTITCRASQSIGSSLH
WYQQKPDQSPKLLIKYGSQSISGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQSSISPRRTFGQG
TKVEIK
J693M2S2R-72Vk
432
EIVLTQSPDFQSVTPKEKVTITCRASQTIGRSLH
WYQQKPDQSPKLLIKYASQSISGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQRKSSPTPTFGQG
TKVEIK
J693M2S2R-75Vk
433
EIVLTQSPDFQSVTPKEKVTITCRASQRIGRQLH
WYQQKPDQSPKLLIKHPSQSISGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQSSSLPPQTFGQG
TKVEIK
J693M2S2R-77Vk
434
EIVLTQSPDFQSVTPKEKVTITCRASQSIGSSLH
WYQQKPDQSPKLLIKHTSQSISGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQICRSPSPTFGQG
TKVEIK
J693M2S2R-78Vk
435
EIVLTQSPDFQSVTPKEKVTITCRASQSIGSSLH
WYQQKPDQSPKLLIKYASQSSSGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQSSGSPAPTFGQG
TKVEIK
J693M2S2R-79Vk
436
EIVLTQSPDFQSVTPKEKVTITCRASQSIGSSLH
WYQQKPDQSPKLLIKYSSQSTSGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQISSSPPPTFGQG
TKVEIK
J693M2S2R-7Vk
437
EIVLTQSPDFQSVTPKEKVTITCRASQTIGNSLH
WYQQKPDQSPKLLIKHASQSNSGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQTMTSPPPTFGQG
TKVEIK
J693M2S2R-80Vk
438
EIVLTQSPDFQSVTPKEKVTITCRASQSIGSSLH
WYQQKPDQSPKLLIKHASQSISGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQSRSSPSPTFGQG
TKVEIK
J693M2S2R-81Vk
439
EIVLTQSPDFQSVTPKEKVTITCRASQSIGSSLH
WYQQKPDQSPKLLIKHASQSISGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQRRWSPPPTFGQG
TKVEIK
J693M2S2R-82Vk
440
EIVLTQSPDFQSVTPKEKVTITCRASQSIGSSLH
WYQQKPDQSPKLLIKYASQSNSGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQISCLPLPTFGQG
TKVEIK
J693M2S2R-83Vk
441
EIVLTQSPDFQSVTPKEKVTITCRASQRIGSSLH
WYQQKPDQSPKLLIKHASQSISGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQSISLPPPTFGQG
TKVEIK
J693M2S2R-84Vk
442
EIVLTQSPDFQSVTPKEKVTITCRASQSIGRNLH
WYQQKPDQSPKLLIKHTSQSLSGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQTSTLPPQTFGQG
TKVEIK
J693M2S2R-85Vk
443
EIVLTQSPDFQSVTPKEKVTITCRASQSIGRSLH
WYQQKPDQSPKLLIKYASQSISGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQSRNSPQPTFGQG
TKVEIK
J693M2S2R-86Vk
444
EIVLTQSPDFQSVTPKEKVTITCRASQSIGTRLH
WYQQKPDQSPKLLIKYVSQSISGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQSSHSPPPTFGQG
TKVEIK
J693M2S2R-87Vk
445
EIVLTQSPDFQSVTPKEKVTITCRASQSIGSCLH
WYQQKPDQSPKLLIKHRSQSISGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQWSSSPPPTFGQG
TKVEIK
J693M2S2R-89Vk
446
EIVLTQSPDFQSVTPKEKVTITCRASQRIGSSLH
WYQQKPDQSPKLLIKHPSQSISGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQTSGSPSHTFGQG
TKVEIK
J693M2S2R-8Vk
447
EIVLTQSPDFQSVTPKEKVTITCRASQGIGSSLH
WYQQKPDQSPKLLIKYESQSISGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQSSSSPPPTFGQG
TKVEIK
J693M2S2R-90Vk
448
EIVLTQSPDFQSVTPKEKVTITCRASQSIGSSLH
WYQQKPDQSPKLLIKHDSQSISGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQGSSSPPTTFGQG
TKVEIK
J693M2S2R-91Vk
449
EIVLTQSPDFQSVTPKEKVTITCRASQTIGSNLH
WYQQKPDQSPKLLIKHASQSISGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQRISSPPSTFGQG
TKVEIK
J693M2S2R-92Vk
450
EIVLTQSPDFQSVTPKEKVTITCRASQTIGSSLH
WYQQKPDQSPKLLIKHASQSVSGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQSCSSPPSTFGQG
TKVEIK
J693M2S2R-93Vk
451
EIVLTQSPDFQSVTPKEKVTITCRASQTIGSSLH
WYQQKPDQSPKLLIKYVSQSLSGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQTISSPLPTFGQG
TKVEIK
J693M2S2R-95Vk
452
EIVLTQSPDFQSVTPKEKVTITCRASQSIGSRLH
WYQQKPDQSPKLLIKHASQSISGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQSSCSPAHTFGQG
TKVEIK
J703M1S3-11Vk
453
EIVLTQSPDFQSVTPKEKVTITCRDSRCIGSNLH
WYQQKPDQSPKLLIKHASQSSSGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQCSSSPPPTFGQG
TKVEIK
J703M1S3-13Vk
454
EIVLTQSPDFQSVTPKEKVTITCRASQSIGSTLH
WYQQKPDQSPKLLIKHASQSNSGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQSSSLPPPTFGQG
TKVEIK
J703M1S3-16Vk
455
EIVLTQSPDFQSVTPKEKVTITCRASQSIGDSLH
WYQQKPDQSPKLLIKHASQSKSGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQGSTSPPRTFGQG
TKVEIK
J703M1S3-19Vk
456
EIVLTQSPDFQSVTPKEKVTITCRASQSIGSSLH
WYQQKPDQSPKLLIKHGSQSSSGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQSWSSPIPTFGQG
TKVEIK
J703M1S3-22Vk
457
EIVLTQSPDFQSVTPKEKVTITCRASQSIGSRLH
WYQQKPDQSPKLLIKYASQSTSGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQSSNLPSPTFGQG
TKVEIK
J703M1S3-26Vk
458
EIVLTQSPDFQSVTPKEKVTITCRASQSIGSRLH
WYQQKPDQSPKLLIKHASQSTSGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQSGSSPPRTFGQG
TKVEIK
J703M1S3-29Vk
459
EIVLTQSPDFQSVTPKEKVTITCRASQSIGSSLH
WYQQKPDQSPKLLIKHASQSISGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQRTSSPVRTFGQG
TKVEIK
J703M1S3-2Vk
460
EIVLTQSPDFQSVTPKEKVTITCRASQSIGNTLH
WYQQKPDQSPKLLIKHVSQSVSGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQKVSSPSPTFGQG
TKVEIK
J703M1S3-30Vk
461
EIVLTQSPDFQSVTPKEKVTITCRASQRIGSSLH
WYQQKPDQSPKLLIKHASQSVSGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQSRSSPPPTFGQG
TKVEIK
J703M1S3-33Vk
462
EIVLTQSPDFQSVTPKEKVTITCRASQSIGSSLH
WYQQKPDQSPKLLIKHASQSTSGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQSSSSPPSTFGQG
TKVEIK
J703M1S3-34Vk
463
EIVLTQSPDFQSVTPKEKVTITCRASQSIGSSLH
WYQQKPDQSPKLLIKYASQSISGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQNSSSPSTTFGQG
TKVEIK
J703M1S3-57Vk
464
EIVLTQSPDFQSVTPKEKVTITCRASQCIGSSLH
WYQQKPDQSPKLLIKHESQSSSGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQRCTSPSPTFGQG
TKVEIK
J703M1S3-5Vk
465
EIVLTQSPDFQSVTPKEKVTITCRASQRIGSSLH
WYQQKPDQSPKLLIKHPSQSDSGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQNCSLPLPTFGQG
TKVEIK
J703M1S3-62Vk
466
EIVLTQSPDFQSVTPKEKVTITCRASQCIGSSLH
WYQQKPDQSPKLLIKHASQSTSGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQGISSPPQTFGQG
TKVEIK
J703M1S3-69Vk
467
EIVLTQSPDFQSVTPKEKVTITCRASQSIGSSLH
WYQQKPDQSPKLLIKHVSQSLSGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQRSSSPSPTFGQG
TKVEIK
J703M1S3-71Vk
468
EIVLTQSPDFQSVTPKEKVTITCRASQSIGSSLH
WYQQKPDQSPKLLIKHPSQSLSGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQSIRLPPSTFGQG
TKVEIK
J703M1S3-78Vk
469
EIVLTQSPDFQSVTPKEKVTITCRANQSIGGSLH
WYQQKPDQSPKLLIKHASQSKSGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQDSRSPTRTFGQG
TKVEIK
J703M1S3-79Vk
470
EIVLTQSPDFQSVTPKEKVTITCRASQSIGSGLH
WYQQKPDQSPKLLIKHTSQSISGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQSSSLPHPTFGQG
TKVEIK
J703M1S3-7Vk
471
EIVLTQSPDFQSVTPKEKVTITCRASQSIGSSLH
WYQQKPDQSPKLLIKHASQSTSGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQSSSSPTPTFGQG
TKVEIK
J703M1S3-81Vk
472
EIVLTQSPDFQSVTPKEKVTITCRASQSIGSRLH
WYQQKPDQSPKLLIKYPSQSRSGVPSRFSGSGSG
TDLTLTINSLEAEDAATYYCHQNGSLPPPTFGQG
TKVEIK
J703M1S3-82Vk
473
EIVLTQSPDFQSVTPKEKVTITCRASQSIGSSLH
WYQQKPDQSPKLLIKHASQSISGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQNSSSPPPTFGQG
TKVEIK
J703M1S3-86Vk
474
EIVLTQSPDFQSVTPKEKVTITCRASQSIGSALH
WYQQKPDQSPKLLIKHASQSLSGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQSSILPRPTFGQG
TKVEIK
J703M1S3-90Vk
475
EIVLTQSPDFQSVTPKEKVTITCRASQSIGSNLH
WYQQKPDQSPKLLIKHASQSISGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQTRTSPPLTFGQG
TKVEIK
J703M1S3-93Vk
476
EIVLTQSPDFQSVTPKEKVTITCRASQKIGSSLH
WYQQKPDQSPKLLIKYGSQSTSGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQCISLPTPTFGQG
TKVEIK
J703M1S3-94Vk
477
EIVLTQSPDFQSVTPKEKVAITCRASQRIGSSLH
WYQQKPDQSPKLLIKYASQSISGVPSRFSGSGSG
TDFTLTINSLEAEDAATYYCHQNSSLPPPTFGQG
TKVEIK
TABLE 13
Amino acid residues observed in affinity matured AE11-5 antibodies
AE11-5 Heavy chain variable region (SEQ ID NO: 1073)
AE11-5VH
1234567890123456789012345678901234567890123456789012a345678901
EVQLVQSGAEVKKPGSSAKVSCKASGGTFSSYAISWVRQAPGQGLEWMGGIIPILGTANYAQ
V NW TTT WT FRSPI
TY SV M TDAST
GI P L I NGS
AN G V P V
F N I H
R V A
L K R
F M
L
234567890123456789012abc345678901234567890abc1234567890123
KFLGRVTITADESTSTVYMELSSLRSEDTAVYYCARGLYYDPTRADYWGQGTLVTVSS
Q A SVFFNTSWF
WIVVEFASM
TFP TRKP
ARH IGRA
Q ADI
Y
V
P
N
G
AE11-5 Light chain variable region (SEQ ID NO: 1074)
AE11-5VL
1234567890123456789012345678901234567890123456789012345678901
DIVMTQSPDFHSVTPKEKVTITCRASQSIGSSLHWYQQKPDQSPKLLIRHASQSISGVPSR
E L Q R RR KYV L
T TT P V
N GN T T
I NC G S
C KG S M
G CI E N
K HK D K
Y VM F
W PL R
LY
P
V
2345678901234567890123456789012345a67890123456a
FSGSGSGTDFTLTIHSLEAEDAATYYCHQSSSSPPPTFGQGTQVEIK
N RRRL LS K
NGI AR
GIC SL
TCG RT
CNN TA
ITT QQ
MK HH
V
M
TABLE 14
Individual VH sequences from converted clones
Protein
Sequence
region
SEQ ID NO:
123456789012345678901234567890
J703M1S3
478
EVQLVQSGAEVKKPGSSVKVSCKASGGTFS
#2
WYATSWVRQAPGQGLEWMGGITPILGSPIY
VH
AQKFQGRVTITADESTSTVYMELSSLRSED
TAVYYCARGVYYDHRRADYWGQGTLVTVSS
J703M1S3
CDR-H1
Residues 31-35
WYATS
#2
of SEQ ID
VH
NO.: 478
J703M1S3
CDR-H2
Residues 50-66
GITPILGSPIYAQKFQG
#2
of SEQ ID
VH
NO.: 478
J703M1S3
CDR-H3
Residues 99-109
GVYYDHRRADY
#2
of SEQ ID
VH
NO.: 478
J703M1S3
479
EVQLVQSGAEVKKPGSSVKVSCKASGGTFS
#13
WYAISWVRQAPGQGLEWMGGITPILGAANY
VH
AQKFQGRVTITADESTSTVYMELSSLRSED
TAVYYCARGVYYDPKRADYWGQGTLVTVSS
J703M1S3
CDR-H1
Residues 31-35
WYAIS
#13
of SEQ ID
NO.: 479
J703M1S3
CDR-H2
Residues 50-66
GITPILGAANYAQKFQG
#13
of SEQ ID
VH
NO.: 479
J703M1S3
CDR-H3
Residues 99-109
GVYYDPKRADY
#13
of SEQ ID
VH
NO.: 479
J703M1S3
480
EVQLVQSGAEVKKPGSSVKVSCKASGGTFS
#26
WYAISWVRQAPGQGLEWMGGITPILGTANY
VH
AQKFQGRVTITADESTSTVYMELSSLRSED
TAVYYCARGVYYDPKRADYWGQGTLVTVSS
J703M1S3
CDR-H1
Residues 31-35
WYAIS
#26
of SEQ ID
VH
NO.: 480
J703M1S3
CDR-H2
Residues 50-66
GITPILGTANYAQKFQG
#26
of SEQ ID
VH
NO.: 480
J703M1S3
CDR-H3
Residues 99-109
GVYYDPKRADY
#26
of SEQ ID
VH
NO.: 480
J703M1S3
481
EVQLVQSGAEVKKPGSSVKVSCKASGGTFS
#30
WYAISWVRQAPGQGLEWMGGITPILGSPIY
VH
AQKFQGRVTITADESTSTVYMELSSLRSED
TAVYYCARGVYYDPKRADYWGQGTLVTVSS
J703M1S3
CDR-H1
Residues 31-35
WYAIS
#30
of SEQ ID
VH
NO.: 481
J703M1S3
CDR-H2
Residues 50-66
GITPILGSPIYAQKFQG
#30
of SEQ ID
VH
NO.: 481
J703M1S3
CDR-H3
Residues 99-109
GVYYDPKRADY
#30
of SEQ ID
VH
NO.: 481
J703M1S3
482
EVQLVQSGAEVKKPGSSVKVSCKASGGTFS
#33
WYPISWVRQAPGQGLEWMGGITPILGAGIY
VH
AQKFQGRVTITADESTSTVYMELSSLRSED
TAVYYCARGVYYDFKRADYWGQGTLVTVSS
J703M1S3
CDR-H1
Residues 31-35
WYPIS
#33
of SEQ ID
VH
NO.: 482
J703M1S3
CDR-H2
Residues 50-66
GITPILGAGIYAQKFQG
#33
of SEQ ID
VH
NO.: 482
J703M1S3
CDR-H3
Residues 99-109
GVYYDFKRADY
#33
of SEQ ID
VH
NO.: 482
J703M1S3
483
EVQLVQSGAEVKKPGSSVKVSCKASGGTFS
#35
WYAISWVRQAPGQGLEWMGGITPILGSATY
VH
AQKFQGRVTITADESTSTVYMELSSLRSED
TAVYYCARGIYYDPKRADYWGQGTLVTVSS
J703M1S3
CDR-H1
Residues 31-35
WYAIS
#35
of SEQ ID
VH
NO.: 483
J703M1S3
CDR-H2
Residues 50-66
GITPILGSATYAQKFQG
#35
of SEQ ID
VH
NO.: 483
J703M1S3
CDR-H3
Residues 99-109
GIYYDPKRADY
#35
of SEQ ID
VH
NO.: 483
J703M1S3
484
EVQLVQSGAEVKKPGSSVKVSCKASGGTFS
#38
WYAISWVRQAPGQGLEWMGGITPILGTPIY
VH
AQKFQGRVTITADESTSTVYMELSSLRSED
TAVYYCARGVYYDFKRADYWGQGTLVTVSS
J703M1S3
CDR-H1
Residues 31-35
WYAIS
#38
of SEQ ID
VH
NO.: 484
J703M1S3
CDR-H2
Residues 50-66
GITPILGTPIYAQKFQG
#38
of SEQ ID
VH
NO.: 484
J703M1S3
CDR-H3
Residues 99-109
GVYYDFKRADY
#38
of SEQ ID
VH
NO.: 484
J703M1S3
485
EVQLVQSGAEVKKPGSSVKVSCKASGGTFS
#69
WYAISWVRQAPGQGLEWMGGITPILGSPIY
VH
AQKFQGRVTITADESTSTVYMELSSLRSED
TAVYYCARGIYYDPKRADYWGQGTLVTVSS
J703M1S3
CDR-H1
Residues 31-35
WYAIS
#69
of SEQ ID
VH
NO.: 485
J703M1S3
CDR-H2
Residues 50-66
GITPILGSPIYAQKFQG
#69
of SEQ ID
VH
NO.: 485
J703M1S3
CDR-H3
Residues 99-109
GIYYDPKRADY
#69
of SEQ ID
VH
NO.: 485
J703M1S3
486
EVQLVQSGAEVKKPGSSVKVSCKASGGTFS
#90
WYAISWVRQAPGQGLEWMGGITPILGSPIY
VH
AQKFQGRVTITADESTSTVYMELSSLRSED
TAVYYCARGVYYDYKRADYWGQGTLVTVSS
J703M1S3
CDR-H1
Residues 31-35
WYAIS
#90
of SEQ ID
VH
NO.: 486
J703M1S3
CDR-H2
Residues 50-66
GITPILGSPIYAQKFQG
#90
of SEQ ID
VH
NO.: 486
J703M1S3
CDR-H3
Residues 99-109
GVYYDYKRADY
#90
of SEQ ID
VH
NO.: 486
TABLE 15
Individual clones VL sequences
Protein
Sequence
region
123456789012345678901234567890
J703M1S3
487
EIVLTQSPDFQSVTPKEKVTITCRASQSIG
#2
NTLHWYQQKPDQSPKLLIKHVSQSVSGVPS
VL
RFSGSGSGTDFTLTINSLEAEDAATYYCHQ
KVSSPSPTFGQGTKVEIK
J703M1S3
CDR-L1
Residues 24-34
RASQSIGNTLH
#2
of SEQ ID
VL
NO.: 487
J703M1S3
CDR-L2
Residues 50-56
HVSQSVS
#2
of SEQ ID
VL
NO.: 487
J703M1S3
CDR-L3
Residues 89-98
HQKVSSPSPT
#2
of SEQ
VL
ID NO.: 487
J703M1S3
488
EIVLTQSPDFQSVTPKEKVTITCRASQSIG
#13
STLHWYQQKPDQSPKLLIKHASQSNSGVPS
VL
RFSGSGSGTDFTLTINSLEAEDAATYYCHQ
SSSLPPPTFGQGTKVEI
J703M1S3
CDR-L1
Residues 24-34
RASQSIGSTLH
#13
of SEQ ID
VL
NO.: 488
J703M1S3
CDR-L2
Residues 50-56
HASQSNS
#13
of SEQ ID
VL
NO.: 488
J703M1S3
CDR-L3
Residues 89-98
HQSSSLPPPT
#13
of SEQ
VL
ID NO.: 488
J703M1S3
489
EIVLTQSPDFQSVTPKEKVTITCRASQSIG
#26
SRLHWYQQKPDQSPKLLIKHASQSTSGVPS
VL
RFSGSGSGTDFTLTINSLEAEDAATYYCHQ
SGSSPPRTFGQGTKVEIK
J703M1S3
CDR-L1
Residues 24-34
RASQSIGSRLH
#26
of SEQ ID
VL
NO.: 489
J703M1S3
CDR-L2
Residues 50-56
HASQSTS
#26
of SEQ ID
VL
NO.: 489
J703M1S3
CDR-L3
Residues 89-98
HQSGSSPPRT
#26
of SEQ
VL
ID NO.: 489
J703M1S3
490
EIVLTQSPDFQSVTPKEKVTITCRASQRIG
#30
SSLHWYQQKPDQSPKLLIKHASQSVSGVPS
VL
RFSGSGSGTDFTLTINSLEAEDAATYYCHQ
SRSSPPPTFGQGTKVEIK
J703M1S3
CDR-L1
Residues 24-34
RASQRIGSSLH
#30
of SEQ ID
VL
NO.: 490
J703M1S3
CDR-L2
Residues 50-56
HASQSVS
#30
of SEQ ID
VL
NO.: 490
J703M1S3
CDR-L3
Residues 89-98
HQSRSSPPPT
#30
of SEQ
VL
ID NO.: 490
J703M1S3
491
EIVLTQSPDFQSVTPKEKVTITCRASQSIG
#33
SSLHWYQQKPDQSPKLLIKHASQSTSGVPS
VL
RFSGSGSGTDFTLTINSLEAEDAATYYCHQ
SSSSPPSTFGQGTKVEIK
J703M1S3
CDR-L1
Residues 24-34
RASQSIGSSLH
#33
of SEQ ID
VL
NO.: 491
J703M1S3
CDR-L2
Residues 50-56
HASQSTS
#33
of SEQ ID
VL
NO.: 491
J703M1S3
CDR-L3
Residues 89-98
HQSSSSPPST
#33
of SEQ
VL
ID NO.: 491
J703M1S3
492
EIVLTQSPDFQSVTPKEKVTITCRASQTIG
#35
SSLHWYQQKPDQSPKLLIKHASQSISGVPS
VL
RFSGSGSGTDFTLTINSLEAEDAATYYCHQ
TSSLPTPTFGQGTKVEIK
J703M1S3
CDR-L1
Residues 24-34
RASQTIGSSLH
#35
of SEQ ID
VL
NO.: 492
J703M1S3
CDR-L2
Residues 50-56
HASQSIS
#35
of SEQ ID
VL
NO.: 492
J703M1S3
CDR-L3
Residues 89-98
HQTSSLPTPT
#35
of SEQ
VL
ID NO.: 492
J703M1S3
493
EIVLTQSPDFQSVTPKEKVTITCRASQTIG
#38
SSLHWYQQKPDQSPKLLIKHASQSISGVPS
VL
RFSGSGSGTDFTLTINSLEAEDAATYYCHQ
SSSSPPPTFGQGTKVEIK
J703M1S3
CDR-L1
Residues 24-34
RASQTIGSSLH
#38
of SEQ ID
VL
NO.: 493
J703M1S3
CDR-L2
Residues 50-56
HASQSIS
#38
of SEQ ID
VL
NO.: 493
J703M1S3
CDR-L3
Residues 89-98
HQSSSSPPPT
#38
of SEQ
VL
ID NO.: 493
J703M1S3
494
EIVLTQSPDFQSVTPKEKVTITCRASQSIG
#69
SSLHWYQQKPDQSPKLLIKHVSQSLSGVPS
VL
RFSGSGSGTDFTLTINSLEAEDAATYYCHQ
RSSSPSPTFGQGTKVEIK
J703M1S3
CDR-L1
Residues 24-34
RASQSIGSSLH
#69
of SEQ ID
VL
NO.: 494
J703M1S3
CDR-L2
Residues 50-56
HVSQSLS
#69
of SEQ ID
VL
NO.: 494
J703M1S3
CDR-L3
Residues 89-98
HQRSSSPSPT
#69
of SEQ
VL
ID NO.: 494
J703M1S3
495
EIVLTQSPDFQSVTPKEKVTITCRASQSIG
#90
SNLHWYQQKPDQSPKLLIKHASQSISGVPS
VL
RFSGSGSGTDFTLTINSLEAEDAATYYCHQ
TRTSPPLTFGQGTKVEIK
J703M1S3
CDR-L1
Residues 24-34
RASQSIGSNLH
#90
of SEQ ID
VL
NO.: 495
J703M1S3
CDR-L2
Residues 50-56
HASQSIS
#90
of SEQ ID
VL
NO.: 495
J703M1S3
CDR-L3
Residues 89-98
HQTRTSPPLT
#90
of SEQ
VL
ID NO.: 495
TABLE 16
AE11-5 affinity matured scFv clones converted to full length IgG
Full length
ScFv
IgG (protein)
clone name
HC plasmid
LC plasmid
name
J703M1S3#2
pJP368; pHybE-hCg1,z,non-
pJP369; pHybE-hCk V3-
AE11-5 AM1
a,mut(234,235)-J703M1S3#2
J703M1S31#2
J703M1S3#13
pJP370; pHybE-hCg1,z,non-
pJP371; pHybE-hCk V3-
AE11-5 AM2
a,mut(234,235)-J703M1S3#13
J703M1S3#13
J703M1S3#26
pJP372; pHybE-hCg1,z,non-
pJP373; pHybE-hCk V3-
AE11-5 AM3
a,mut(234,235)-J703M1S3#26
J703M1S3#26
J703M1S3#30
pJP374; pHybE-hCg1,z,non-
pJP375; pHybE-hCk V3-
AE11-5 AM4
a,mut(234,235)-J703M1S3#30
J703M1S3#30
J703M1S3#33
pJP376; pHybE-hCg1,z,non-
pJP377; pHybE-hCk V3-
AE11-5 AM5
a,mut(234,235)-J703M1S3#33
J703M1S3#33
J703M1S3#35
pJP378; pHybE-hCg1,z,non-
pJP379; pHybE-hCk V3-
AE11-5 AM6
a,mut(234,235)-J703M1S3#35
J703M1S3#35
J703M1S3#38
pJP382; pHybE-hCg1,z,non-
pJP383; pHybE-hCk V3-
AE11-5 AM8
a,mut(234,235)-J703M1S3#38
J703M1S3#38
J703M1S3#69
pJP384; pHybE-hCg1,z,non-
pJP385; pHybE-hCk V3-
AE11-5 AM9
a,mut(234,235)-J703M1S3#69
J703M1S3#69
J703M1S3#90
pJP386; pHybE-hCg1,z,non-
pJP387; pHybE-hCk V3-
AE11-5 AM10
a,mut(234,235)-J703M1S3#90
J703M1S3#90
The following protocol is used to characterize the binding of TNF antibodies to biotinylated human or cyno TNF by enzyme-linked immunosorbent assay (ELISA). An ELISA plate was coated with 50 μl per well of goat anti human IgG-Fc at 2 μg/ml, overnight at 4° C. The plate was washed 3 times with PBS/Tween. 50 μl Mab diluted to 1 μg/ml in PBS/0.1% BSA was added to appropriate wells and incubated for 1 hour at room temperature (RT). The plate was washed 3 times with PBS/Tween. 50 μl of serial diluted biotin-human TNF was added to appropriate wells and incubated for 1 hour at RT. The plate was washed 3 times with PBS/Tween. 50 μl of streptavidin-HRP diluted 1:10,000 in PBS/0.1% BSA was added to appropriate wells and incubated for 1 hour at RT. The plate was washed 3 times with PBS/Tween. 50 μl of TMB was added to appropriate wells and the reaction was allowed to proceed for 1 minute. The reaction was stopped with 50 μl/well 2N H2SO4 and the absorbance read at 450 nm. Results are shown in Table 17.
TABLE 17
EC50 in hTNF
EC50 in cynoTNF
IgG Name
ELISA (nM)
ELISA (nM)
AE11-5-AM1
1.06
2.14
AE11-5-AM2
522.5
>845
AE11-5-AM3
1.57
1.55
AE11-5-AM4
18.32
750.3
AE11-5-AM5
17.7
2.2
AE11-5-AM6
1.37
>720
AE11-5-AM7
10.32
1.26
AE11-5-AM8
250.2
58.58
AE11-5-AM9
16.72
5.29
AE11-5-AM10
0.98
0.28
Human TNF was prepared at Abbott Bioresearch Center (Worcester, Mass., US) and received from the Biologics Pharmacy. Mouse TNF was prepared at Abbott Bioresearch Center and received from the Biologics Pharmacy. Rat TNF was prepared at Abbott Bioresearch Center and received from the Biologics Pharmacy. Rabbit TNF was purchased from R&D Systems. Rhesus/Macaque TNF (rhTNF) was purchased from R&D Systems. Actinomycin was purchased from Sigma Aldrich and resuspended at a stock concentration of 10 mg/mL in DMSO.
Assay Media: 10% FBS (Hyclone #SH30070.03), Gibco reagents: RPMI 1640 (#21870), 2 mM L-glutamine (#25030), 50 units/mL penicillin/50 μg/mL streptomycin (#15140), 0.1 mM MEM non-essential amino acids (#11140) and 5.5×10−5 M 2-mercaptoethanol (#21985-023).
L929 cells were grown to a semi-confluent density and harvested using 0.05% tryspin (Gibco #25300). The cells were washed with PBS, counted, and resuspended at 1E6 cells/mL in assay media containing 4 μg/mL actinomycin D. The cells were seeded in a 96-well plate (Costar #3599) at a volume of 50 μL and 5E4 cells/well. Wells received 50 μL of assay media, bringing the volume to 100 μL.
A test sample was prepared as follows. The test and control IgG proteins were diluted to a 4× concentration in assay media and serial 1:3 dilutions were performed. TNF species were diluted to the following concentrations in assay media: 400 pg/mL huTNF, 200 pg/mL muTNF, 600 pg/mL ratTNF, and 100 pg/mL rabTNF. Antibody sample (200 μL) was added to the TNF (200 μL) in a 1:2 dilution scheme and allowed to incubate for 0.5 hour at room temperature.
To measure huTNF neutralization potency in this assay, the antibody/TNF solution was added to the plated cells at 100 μL for a final concentration at 375 nM-0.019 nM. The final concentration of TNF was as follows: 100 pg/mL huTNF, 50 pg/mL muTNF, 150 pg/mL ratTNF, and 25 pg/mL rabTNF. The plates were incubated for 20 hours at 37° C., 5% CO2. To quantitate viability, 100 μL was removed from the wells and 10 μL of WST-1 reagent (Roche cat #11644807001) was added. Plates were incubated under assay conditions for 3.5 hours, centrifuged at 500×g, and 75 μL of supernatant transferred to an ELISA plate (Costar cat #3369). The plates were read at OD 420-600 nm on a Spectromax 190 ELISA plate reader. The neutralization potency of selected TNF/IL-17 DVD-Ig binding proteins is shown in Table 18.
TABLE 18
hu TNF neutralization
rhesus TNF neutralization IC50
IgG Name
IC50 (nM)
(nM)
AE11-5 AM1
0.439
0.251
AE11-5 AM2
1.241
0.756
AE11-5 AM3
0.291
0.165
AE11-5 AM4
0.259
0.109
AE11-5 AM5
0.968
0.613
AE11-5 AM6
2.029
0.652
AE11-5 AM7
0.049
0.104
AE11-5 AM8
1.356
3.040
AE11-5 AM9
0.391
0.123
AE11-5 AM10
0.678
0.140
The mouse anti-human TNF antibody MAK-195 was humanized and affinity-matured to generate a panel of humanized MAK195 variants that have cross-reactivity to cyno-TNF and improved affinity and binding kinetics against both human and cyno TNF.
To improve the affinity of hMAK195 to TNF, hypermutated CDR residues were identified from other human antibody sequences in the IgBLAST database that also shared high identity to germlines VH3-53 and IGKV1-39. The corresponding hMAK195 CDR residues were then subjected to limited mutagenesis by PCR with primers having low degeneracy at these positions to create three antibody libraries in the scFv format. The first library contained mutations at residues 31, 32, 33, 35, 50, 52, 53, 54, 56 and 58 in the VH CDR1 and 2 (Kabat numbering); the second library at residues 95 to 100, 100a, 101, and 102 in VH CDR3; and the third library at residues 28, 30, 31, 32, 50, 53, 92, 93, 94, and 95 in the three VL CDRs. To further increase the identity of hMAK195 to the human germline framework sequences, a binary degeneracy at VH positions 60 (D/A), 61 (S/D), 62 (T/S), 63 (L/V), and 65 (S/G) were introduced into the first library. Also, a binary degeneracy at VL positions 24 (K/R), 33 (V/L), 54 (R/L), 55 (H/Q), 56 (T/S), 91 (H/S) and 96 (F/Y) were introduced into the third library.
These hMAK195 variants were selected against a low concentration of biotinylated TNF for improved on-rate, off-rate, or both were carried out and antibody protein sequences of affinity-modulated hMAK195 were recovered for converting back to IgG for further characterization. All three libraries were selected separately for the ability to bind human or cynomolgus monkey TNF in the presence of decreasing concentrations of biotinylated human or cynomolgus monkey TNF antigens. All mutated CDR sequences recovered from library selections were recombined into additional libraries and the recombined libraries were subjected to more stringent selection conditions before individual antibodies are identified.
Table 19 provides a list of amino acid sequences of VH and VL of the humanized MAK-195 which were subjected to the affinity maturation selection protocol Amino acid residues of individual CDRs of each VH and VL sequence are indicated in bold.
TABLE 19
List of amino acid sequences of affinity matured hMAK195
VH variants
SEQ ID
Clone
NO:
VH
rHC1_B8
496
EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYGVSWVRQAPGK
GLEWVSIIRGDGSTDYASTLKSRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWQHGPVAYWGQGTLVTVSS
rHC1_H12
497
EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYGVSWVRQAPGK
GLEWVSIIRGDGSTDYADSVKSRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWHHGPVAYWGQGTLVTVSS
H1 + H2_E1
498
EVQLVESGGGLVQPGGSLRLSCAASGFTFSAYGVNWVRQAPGK
GLEWVSIIWGDGATDYADSVKGRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWHHGPVAYWGQGTLVTVSS
rHC1_A2
499
EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYGVNWVRQAPGK
GLEWVSMISSDGFTDYASSVKSRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWQHGPVAYWGQGTLVTVSS
rHC1_H6
500
EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYGVTWVRQAPGK
GLEWVSMIAADGSTDYASSVKGRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWQHGPVAYWGQGTLVTVSS
H1 + H2_D7
501
EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYGVTWVRQAPGK
GLEWVSMIRADGSTDYASSLKSRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWHHGPVAYWGQGTLVTVSS
H1 + H2_D9
502
EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYGVTWVRQAPGK
GLEWVSMIRDDGSTDYADTVKGRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWHHGPVAYWGQGTLVTVSS
rHC1_A10
503
EVQLVESGGGLVQPGGSLRLSCAASGETFSHIGVSWVRQAPGK
GLEWVSMISYAGSTDYASTVKGRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCARLLHKGPIDYWGQGTLVTVSS
H1 + H2_A5
504
EVQLVESGGGLVQPGGSLRLSCAASGFTFSDFGVNWVRQAPGK
GLEWVSMIWSDGSTDYADTVKSRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWHHGPVAYWGQGTLVTVSS
H1 + H2_F8
505
EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYGVSWVRQAPGK
GLEWVSIIRADGSTDYASSLKGRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWHHGPVAYWGQGTLVTVSS
rHC1_D1
506
EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYGVQWVRQAPGK
GLEWVSMIRGDGSTDYASSLKGRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCARPSHHGLIDNWGQGTLVTVSS
rHC2_C2
507
EVQLVESGGGLVQPGGSLRLSCAASGFTFSELGVNWVRQAPGK
GLEWVSYISDVGSTYYASTVKGRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCARDWHHGRFDYWGQGTLVTVSS
rHC1_G4
508
EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYGVSWVRQAPGK
GLEWVSLIRADGSTDYADSLKGRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWHHGPVAYWGQGTLVTVSS
rHC1_F3
509
EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYGVTWVRQAPGK
GLEWVSMIRADGFTDYADSVKGRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCARDWQHGPSVYWGQGTLVTVSS
rHC1_B4
510
EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYGVTWVRQAPGK
GLEWVSIIRADGVTDYASSVKGRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWHHGPVAYWGQGTLVTVSS
H1 + H2_G3
511
EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYGVHWVRQAPGK
GLEWVSMIGADGYTDYADSLKGRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWHHGPVAYWGQGTLVTVSS
rHC1_D7
512
EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYGVSWVRQAPGK
GLEWVSMISADGSTDYASSVKGRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWHHGPVAYWGQGTLVTVSS
rHC1_D5
513
EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYGVTWVRQAPGK
GLEWVSMIRSDGFTDYADSVKGRFTISRDNSKNTLYLQMNSLR
TEDTAVYYCAREWHHGPVAYWGQGTLVTVSS
H1 + H2_E4
514
EVQLVESGGGLVQPGGSLRLSCAASGFTFSEYGVNWVRQAPGK
GLEWVSIIWHDGSTAYADTVKSRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWHHGPVAYWGQGTLVTVSS
rHC1_E10
515
EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYGVTWVRQAPGK
GLEWVSLIRGDGSTDYASSVKSRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWHHGPVAYWGQGTLVTVSS
H1 + H2_B6
516
EVQLVESGGGLVQPGGSLRLSCAASGFTFSNFGVSWVRQAPGK
GLEWVSMIWGDGSTDYADSLKGRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWHHGPVAYWGQGTLVTVSS
rHC1_B7
517
EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYGVTWVRQAPGK
GLEWVSMIRDDGSTYYASTVKGRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWQHGPLGYWGQGTLVTVSS
H1 + H2_G8
518
EVQLVESGGGLVQPGGSLRLSCAASGFTFSDFGVNWVRQAPGK
GLEWVSMIWAGGSTAYASTVKGRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWHHGPVAYWGQGTLVTVSS
rHC1_G5
519
EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYGVSWVRQAPGK
GLEWVSLIGADGSTDYASTVKGRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWQYGPLAYWGQGTLVTVSS
H1 + H2_F1
520
EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYGVTWVRQAPGK
GLEWVSMIEGDGGTHYASSVKGRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWHHGPVAYWGQGTLVTVSS
rHC19
521
EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYGVTWVRQAPGK
GLEWVSMIWADGSTHYASSVKGRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWQHGPAAYWGQGTLVTVSS
H1 + H2_A10
522
EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYGVNWVRQAXGK
GLEWVSMISADGTTDYASSVKGRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWHHGPVAYWGQGTLVTVSS
rHC1_B9
523
EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYGVTWVRQAPGK
GLEWVSIIRGDGTTDYASTVKGRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWQHGPLGYWGQGTLVTVSS
H1 + H2_F7
524
EVQLVESGGGLVQPGGSLRLSCAASGFTFSHYGVGWVRQAPGK
GLEWVSMIWGAGSTNYADTVKSRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWHHGPVAYWGQGTLVTVSS
H1 + H2_B1
525
EVQLVESGGGLVQPGGSLRLSCAASGFTFSSFGVNWVRQAPGK
GLEWVSMIWADGTTDYADSLKGRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWHHGPVAYWGQGTLVTVSS
rHC1_H9
526
EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYGVTWVRQAPGK
GLEWVSVIGGDGYTDYADSVKGRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWQHGPVAYWGQGTLVTVSS
H1 + H2_A12
527
EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYGVTWVRQAPGX
GLEWVSMISSDGYTDYASSVKGRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWHHGPVAYWGQGTLVTVSS
rHC2_G8
528
EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYGVSWVRQAPGK
GLEWVSMIWSDGSTHYADTVKSRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWHHGPVAYWGQGTLVTVSS
rHC2_B4
529
EVQLVESGGGLVQPGGSLRLSCAASGFTFSQLGVTWVRQAPGK
GLEWVSTISDAGSTYYASSVKGRFTIIRINSKNTLYLQMNSLR
AEDTAVYYCARDWHHGRFAYWGQGTLVTVSS
H1 + H2_G5
530
EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYGVSWVRQAPGK
GLEWVSIIRGDGSTYYASSLKGRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWHHGPVAYWGQGTLVTVSS
rHC1_C6
531
EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYGVQWVRQAPGK
GLEWVSMIRDDGSTSYASTVKGRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWHHGPVAYWGQGTLVTVSS
rHC1_F5
532
EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYGVTWVRQAPGK
GLEWVSIIRGDGSTDYASSVKSRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWQHGPVAYWGQGTLVTVSS
H1 + H2_B4
533
EVQLVESGGGLVQPGGSLRLSCAASGFTFSAYGVNWVRQAPGK
GLEWVSMISGDGSTDYADSVKSRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWHHGPVAYWGQGTLVTVSS
H1 + H2_F6
534
EVQLVESGGGLVQPGGSLRLSCAASGFTFSHFGVTWVRQAPGK
GLEWVSNIWASGSTDYADSVKGRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWHHGPVAYWGQGTLVTVSS
rHC1_B6
535
EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYGVTWVRQAPGK
GLEWVSMIRADGSTDYADSVKGRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWQHGPVGYWGQGTLVTVSS
H1 + H2_A3
536
EVQLVESGGGLVQPGGSLRLSCAASGFTFSNFGVNWVRQAPGK
GLEWVSVIWGDGSTAYASSVKSRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWHHGPVAYWGQGTLVTVSS
rHC1_D10
537
EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYGVSWVRQAPGK
GLEWVSIIRGDGSTDYASSVKGRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWQHGPLAYWGQGTLVTVSS
rHC18
538
EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYGVTWVRQAPGK
GLEWVSMIWSDGSTDYASSVKGRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWQHGPVAYWGQGTLVTVSS
S4-18
539
EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYGVTWVRQAPGK
GLEWVSMIWADGSTHYADSVKSRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWQHGPLAYWGQGTLVTVSS
rHC2_E6
540
EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYGVSWVRQAPGK
GLEWVSLIRGDGSTDYASSLKGRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWHHGPVAYWGQGTLVTVSS
H1 + H2_D4
541
EVQLVESGGGLVQPGGSLRISCAASGFTFSAFGVSWVRQAPGK
GLEWVSMIWGDGSTDYASSVKGRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWHHGPVAYWGQGTLVTVSS
rHC2_F8
542
EVQLVESGGGLVQPGGSLRLSCAASGFTFSDLGVNWVRQAPGK
GLEWVSTISDIGSTYYASTVKGRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCARDWHNGRFDYWGQGTLVTVSS
rHC1_F10
543
EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYGVSWVRQAPGK
GLEWVSIIRGDGFTDYADSVKGRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWHHGPVAYWGQGTLVTVSS
H1 + H2_C12
544
EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYGVTWVRQAPGK
GLEWVSIIRADGSTDYASSVKGRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWHHGPVAYWGQGTLVTVSS
H1 + H2_C11
545
EVQLVESGGGLVQPGGSLRLSCAASGFTFSHFGVNWVRQAPGK
GLEWVSIIWGDGSTAYASSVKGRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWHHGPVAYWGQGTLVTVSS
rHC1_C4
546
EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYGVEWVRQAPGK
GLEWVSKIWADGSTDYADSLKSRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWQHGPLAYWGQGTLVTVSS
H1 + H2_E12
547
EVQLVESGGGLVQPGGSLRLSCAASGFTFSDYGVNWVRQAPGK
GLEWVSLIWGDGTTDYASSVKGRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWHHGPVAYWGQGTLVTVSS
H1 + H2_C4
548
EVQLVESGGGLVQPGGSLRLSCAASGFTFSYFGVSWVRQAPGK
GLEWVSMIWGDGSTDYADSVKGRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWHHGPVAYWGQGTLVTVSS
rHC1_F9
549
EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYGVTWVRQAPGK
GLEWVSMIRSDGSTDYADTLKGRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWQHGPLAYWGQGTLVTVSS
H1 + H2_B5
550
EVQLVESGGGLVQPGGSLRLSCAASGFTFSDFGVNWVRQAPGK
GLEWVSIIWSDGSTDYASSLKGRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWHHGPVAYWGQGTLVTVSS
S4-34
551
EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYGVSWVRQAPGK
GLEWVSMIWADGSTHYADTVKGRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWQHGPLAYWGQGTLVTVSS
H1 + H2_C2
552
EVQLVESGGGLVQPGGSLRLSCAASGFTFSEFGVNWVRQAPGK
GLEWVSMIWGNGATDYASSVKSRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWHHGPVAYWGQGTLVTVSS
H1 + H2_F11
553
EVQLVESGGGLVQPGGSLRLSCAASGFTFSDFGVNWVRQAPGK
GLEWVSMIWGDGTTAYASSVKSRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWHHGPVAYWGQGTLVTVSS
rHC2_E9
554
EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYGVTWVRQAPGK
GLEWVSMIRADGSTDYASSVKGRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWHHGPVAYWGQGTLVTVSS
H1 + H2_B2
555
EVQLVESGGGLVQPGGSLRLSCAASGFTFSDFGVNWVRQAPGK
GLEWVSMIWGDGSTDYADSLKGRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWHHGPVAYWGQGTLVTVSS
H1 + H2_E9
556
EVQLVESGGGLVQPGGSLRLSCAASGFTFSNFGVNWVRQAXGK
GLEWVSMIWGDGSTDYADSLKGRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWHHGPVAYWGQGTLVTVSS
rHC1_A6
557
EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYGVSWVRQAPGK
GLEWVSMIGSDGFTDYASSLKGRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWQHGPVAYWGQGTLVTVSS
H1 + H2_C8
558
EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYGVTWVRQTPGK
GLEWVSMIRGDGSTDYASSVKGRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWHHGPVAYWGQGTLVTVSS
H1 + H2_C5
559
EVQLVESGGGLVQPGGSLRLSCAASGFTFSDFGVSWVRQAPGK
GLEWVSQIWGDGSTDYADSLKGRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWHHGPVAYWGQGTLVTVSS
rHC2_D5
560
EVQLVESGGGLVQPGGSLRLSCAASGFTFSQLGVTWVRQAPGK
GLEWVSTISDAGSTYYASSVKGRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCARDWHHGRFAYWGQGTLVTVSS
rHC1_C7
561
EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYGVTWVRQAPGK
GLEWVSMIRADGSTDYASSVKGRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCARDWQHGPLGYWGQGTLVTVSS
H1 + H2_C3
562
EVQLVESGGGLVQPGGSLRLSCAASGFTFSAYGVHWVRQAPGK
GLEWVSMIWGDGSTDYADSVKSRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWHHGPVAYWGQGTLVTVSS
rHC1_G7
563
EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYGVTWVRQAPGK
GLEWVSMIRGDGTTDYASSVKGRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCARDWQHGPIGYWGQGTLVTVSS
rHC1_A5
564
EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYGVTWVRQAPGK
GLEWVSMIWADGYTDYADSVKGRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWQHGPLAYWGQGTLVTVSS
H1 + H2_G9
565
EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYGVEWVRQAPGK
GLEWVSKIWGDGTTDYADTLKGRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWHHGPVAYWGQGTLVTVSS
H1 + H2_E2
566
EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYGVTWVRQAPGK
GLEWVSMIGGEGRTDYADSVKGRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWHHGPVAYWGQGTLVTVSS
rHC1_C9
567
EVQLVESGGGLVQPGGSLRLSCAASGFTFSNLGVNWVRQAPGK
GLEWVSMIWDVGSTDYASSVKGRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCARDWHHGLFDYWGQGTLVTVSS
rHC1_G6
568
EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYGVTWVRQAPGK
GLEWVSMIMGDGYTDYADSVKGRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWQHGPVAYWGQGTLVTVSS
rHC1_C1
569
EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYGVTWVRQAPGK
GLEWVSMIRDDGATDYADSVKGRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWQHGPLAYWGQGTLVTVSS
rHC1_C2
570
EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYGVTWVRQAPGK
GLEWVSMISGDGYTDYADSVKGRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWQHGPLAYWGQGTLVTVSS
H1 + H2_C1
571
EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYGVTWVRQAPGK
GLEWVSIIRGDGSTDYADSVKGRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWHHGPVAYWGQGTLVTVSS
H1 + H2_B10
572
EVQLVESGGGLVQPGGSLRLSCAASGFTFSNFGVNWVRQAPGX
GLEWVSMIWADGSTDYASTLKGRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWHHGPVAYWGQGTLVTVSS
H1 + H2_E3
573
EVQLVESGGGLVQPGGSLRLSCAASGFTFSAFGVCWVRQAPGK
GLEWVSMIWADGSTDYASSVKGRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWHHGPVAYWGQGTLVTVSS
rHC1_H4
574
EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYGVTWVRQAPGK
GLEWVSMIRSDGSTDYASSVKSRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCARDWQHGPEGYWGQGTLVTVSS
rHC2_A1
575
EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYGVNWVRQAPGK
GLEWVSMIRGDGSTDYASSVKGRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWHHGPVAYWGQGTLVTVSS
H1 + H2_G11
576
EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYGVSWVRQAPGK
GLEWVSLIRSDGSTHYADSLKGRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWHHGPVAYWGQGTLVTVSS
H1 + H2_D8
577
EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYGVNWVRQAPGK
GLEWVSMIRGDGYTDYASSVKGRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWHHGPVAYWGQGTLVTVSS
rHC1_A3
578
EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYGVTWVRQAPGK
GLEWVSMIWADGSTHYADSLKGRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWQHGPVAYWGQGTLVTVSS
S4-31
579
EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYGVQWVRQAPGK
GLEWVSGIGADGSTAYASSLKGRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWQHSGLAYWGQGTLVTVSS
rHC36
580
EVQLVESGGGLVQPGGSLILSCAASGFTFSNYGVSWVRQAPGK
GLEWVSMIWADGSTHYASSLKGRFTISRDNFKNTLYLQMNSLR
AEDTAVYYCAREWQHGPVAYWGQGTLVTVSS
rHC2_G3
581
EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYGVNWVRQAPGK
GLEWVSMIRGDGFTDYASSVKGRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWHHGPVAYWGQGTLVTVSS
rHC1_C10
582
EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYGVSWVRQAPGK
GLEWVSMIAADGSTAYADSLKGRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWHHGPVAYWGQGTLVTVSS
rHC14
583
EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYGVTWVRQAPGK
GLEWVSMIWADGSTHYASSLKGRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWQHGPAAYWGQGTLVTVSS
rHC1_D4
584
EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYGVTWVRQAPGK
GLEWVSMIRGDGSTDYADTVKGRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWHHGPVAYWGQGTLVTVSS
rHC2_D11
585
EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYGVSWVRQAPGK
GLEWVSIISGDGFTDYASSVKGRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWHHGPVAYWGQGTLVTVSS
rHC2_E11
586
EVQLVESGGGLVQPGGSLRLSCAASGFTFSDWGVHWMRQAPGK
GLEWVSTIWDDGSTYYASSVKSRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCARHGHHGPFVYWGQGTLVTVSS
H1 + H2_E7
587
EVQLVESGGGLVQPGGSLRLSCAASXFTFSNFGVNWVRQAPGK
GLEWVSMIWGDGSTDYADSVKSRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWHHGPVAYWGQGTLVTVSS
rHC1_A8
588
EVQLVESGGGLVQPGGSLRLSCAASGFTFSVYGVNWVRQAPGK
GLEWVSMIGDEGSTDYASTVKGRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCARHWHHGAVDYWGQGTLVTVSS
H1 + H2_B9
589
EVQLVESGGGLVQPGGSLRLSCAASGFTFSDYGVNWVRQAPGK
GLEWVSMIWADGSTHYADSLKSRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWHHGPVAYWGQGTLVTVSS
S4-19
590
EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYGVEWVRQAPGK
GLEWVSGIWADGSTHYADTVKSRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWQHGPVAYWGQGTLVTVSS
S4-74
591
EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYGVTWVRQAPGK
GLEWVSMIWADGSTHYADTVKGRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWQHGPLAYWGQGTLVTVSS
rHC1_H2
592
EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYGVTWVRQAPGK
GLEWVSMIRGDGFTDYASSVKGRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWQHGPVAYWGQGTLVTVSS
rHC1_E3
593
EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYGVTWVRQAPGK
GLEWVSMIRADGYTSYASSVKGRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWHHGPVAYWGQGTLVTVSS
rHC34
594
EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYGVTWVRQAPGK
GLEWVSMIWADGSTHYASSVKGRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWQHGPSAYWGQGTLVTVSS
H1 + H2_F2
595
EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYGVNWVRQAPGK
GLEWVSMIRADGSTDYASSVKSRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWHHGPVAYWGQGTLVTVSS
rHC1_D9
596
EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYGVTWVRQAPGK
GLEWVSMIRADGTTDYASSVKSRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWQHGPVGYWGQGTLVTVSS
H1 + H2_E6
597
EVQLVESGGGLVQPGGSLRLSCAASGFTFSNFGVHWVRQAPGK
GLEWVSMIWADGSTVYASSVKSRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWHHGPVAYWGQGTLVTVSS
H1 + H2_F3
598
EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYGVTWVRQAPGK
GLEWVSMIGSDGSTYYADSLKSRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWHHGPVAYWGQGTLVTVSS
rHC1_G11
599
EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYGVTWVRQAPGK
GLEWVSMIRGDGFTDYASSVKGRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWQHGPLGYWGQGTLVTVSS
H1 + H2_D3
600
EVQLVESGGGLVQPGGSLRLSCAASGFTFSDFGVNWVRQAPGK
GLEWVSMIWGDGHTAYASSVKSRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWHHGPVAYWGQGTLVTVSS
H1 + H2_B12
601
EVQLVESGGGLVQPGGSLRLSCAASGFTFSDYGVNWVRQAPGK
GLEWVSMIWAHGATHYASSVKGRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWHHGPVAYWGQGTLVTVSS
H1 + H2_B11
602
EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYGVTWVRQAPGK
GLEWVSLIRDDGSTDYASTLKGRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWHHGPVAYWGQGTLVTVSS
H1 + H2_A8
603
EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYGVTWVRQAPGK
GLEWVSMIWGDGSTDYADSVKSRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWHHGPVAYWGQGTLVTVSS
S4-24
604
EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYGVTWVRQAPGK
GLEWVSMIWADGSTHYASSVKGRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWQHGPVAYWGQGTLVTVSS
rHC1_F11
605
EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYGVSWVRQAPGK
GLEWVSMISADGYTDYADSLKSRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWQHGPVAYWGQGTLVTVSS
H1 + H2_D10
606
EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYGVSWVRQAPGK
GLEWVSMIWADGSTHYASSLKGRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWHHGPVAYWGQGTLVTVSS
rHC2_D6
607
EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYGVNWVRQAPGK
GLEWVSMIGADGYTDYASTVKGRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWHHGPVAYWGQGTLVTVSS
H1 + H2_G4
608
EVQLVESGGGLVQPGGSLRLSCAASGFTFSAFGVSWVRQAPGK
GLEWVSMIWADGSTDYADSVKSRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWHHGPVAYWGQGTLVTVSS
H1 + H2_D11
609
EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYGVTWVRQAPGK
GLEWVSLIRGDGSTDYASSLKGRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWHHGPVAYWGQGTLVTVSS
rHC1_E9
610
EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYGVTWVRQAPGK
GLEWVSMIWADGTTYYADSVKGRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWQHGPLAYWGQGTLVTVSS
rHC1_A12
611
EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYGVQWVRQAPGK
GLEWVSRISGDGSTDYASSVKGRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWHHGPVAYWGQGTLVTVSS
H1 + H2_A2
612
EVQLVESGGGLVQPGGSLRLSCAASGFSFSNFGVNWVRQAPGK
GLEWVSMIWADGSTNYADTVKSRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWHHGPVAYWGQGTLVTVSS
H1 + H2_B7
613
EVQLVESGGGLVQPGGSLRLSCAASGFTFSAYGVSWVRQAPGK
GLEWVSIISADGSTDYASSVKGRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWHHGPVAYWGQGTLVTVSS
rHC1_H8
614
EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYGVTWVRQAPGK
GLEWVSMIRGDGSTDYADSLKGRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWQHGPVAYWGQGTLVTVSS
rHC1_F12
615
EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYGVNWVRQAPGK
GLEWVSMIGADGYTDYADSVKGRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWHHGPVAYWGQGTLVTVSS
rHC1_E5
616
EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYGVTWVRQAPGK
GLEWVSIIRGDGSTDYASTVKGRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWHHGPVAYWGQGTLVTVSS
H1 + H2_A11
617
EVQLVESGGGLVQPGGSLRLSCAASGFTFSDYGVNWVRQAPGK
GLEWVSMIWGSGATDYADSVKSRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWHHGPVAYWGQGTLVTVSS
rHC1_D6
618
EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYGVTWVRQAPGK
GLEWVSMISADGFTDYADSVKGRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWQHGPVAYWGQGTLVTVSS
rHC2_G10
619
EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYGVNWVRQAPGK
GLEWVSMIAADGFTDYASSVKGRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWHHGPVAYWGQGTLVTVSS
rHC1_H3
620
EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYGVTWVRQAPGK
GLEWVSLIAADGSTDYASSVKGRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWQHGPVAYWGQGTLVTVSS
H1 + H2_F10
621
EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYGVTWVRQAPGK
GLEWVSIIRGDGSTAYADTVKGRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWHHGPVAYWGQGTLVTVSS
H1 + H2_C7
622
EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYGVNWVRQAPGK
GLEWVSMIWGDGNTGYASSVKGRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWHHGPVAYWGQGTLVTVSS
rHC1_A9
623
EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYGVTWVRQAPGK
GLEWVSMIRGDGSTDYADSVKGRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWQHGPLAYWGQGTLVTVSS
H1 + H2_E5
624
EVQLVESGGGLVQPGGSLRLSCAASGFTFSDYGVNWVRQAPGK
GLEWVSMIWGDGSTEYADTLKGRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWHHGPVAYWGQGTLVTVSS
rHC62
625
EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYGVSWVRQAPGK
GLEWVSMIWADGSTHYASSLKGRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWQHGPVGYWGQGTLVTVSS
H1 + H2_F4
626
EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYGVYWVRQAPGK
GLEWVSMIWDDGSTEYADSVKSRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWHHGPVAYWGQGTLVTVSS
rHC2_H8
627
EVQLVESGGGLVQPGGSLRLSCAASGFTFSQLGVTWVRQAPGK
GLEWVSTISDAGSTYYASSVKGRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCARDWHHGRFAYWGQGTLVTVSS
rHC2_F4
628
EVQLVESGGGLVQPGGSLRLSCAASGFTFSGPGVNWVRQAPGK
GLEWVSSIWDDGSTYYASSVKSRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCARHSHDGRFDYWGQGTLVTVSS
S4-50
629
EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYGVEWVRQAPGK
GLEWVSGIWADGSTHYADTVKSRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWQHGPVGYWGQGTLVTVSS
H1 + H2_F12
630
EVQLVESGGGLVQPGGSLRLSCAASGFTFSDYGVNWVRQAPGK
GLEWVSMIWGEGSTGYASSVKGRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWHHGPVAYWGQGTLVTVSS
rHC1_E6
631
EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYGVSWVRQAPGK
GLEWVSIIRDDGFTDYASSVKGRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWHHGPVAYWGQGTLVTVSS
rHC1_F2
632
EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYGVTWVRQAPGK
GLEWVSMIGGDGSTDYASSVKGRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWQHGPVAYWGQGTLVTVSS
H1 + H2_G6
633
EVQLVESGGGLVQPGGSLRLSCAASGFTFSDFGVNWVRQAPGK
GLEWVSMIWADGTTDYDDSVKGRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWHHGPVAYWGQGTLVTVSS
rHC2_F5
634
EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYGVNWVRQAPGK
GLEWVSGISADGSTAYDSSLKGRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWHHGPVAYWGQGTLVTVSS
H1 + H2_D6
635
EVQLVESGGGLVQPGGSLRLSCAASGFTFSSYGVSWVRQAPGK
GLEWVSLIRGDGSTYYASTVKGRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWHHGPVAYWGQGTLVTVSS
H1 + H2_A9
636
EVQLVESGGGLVQPGGSLRLSCAASGFTFSNFGVNWVRQAPGK
GLEWVSMIWGDGSTDYASSVKSRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWHHGPVAYWGQGTLVTVSS
H1 + H2_A1
637
EVQLVESGGGLVQPGGSLRLSCAASGFTFSHFGVNWVRQAPGK
GLEWVSMIWADGSTDYASSLKGRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWHHGPVAYWGQGTLVTVSS
rHC60
638
EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYGVSWVRQAPGK
GLEWVSMIWADGSTHYASSLKGRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWQHGPAAYWGQGTLVTVSS
rHC1_C8
639
EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYGVHWVRQAPGK
GLEWVSMIAGDGSTDYASSVKSRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWQHGPVGYWGQGTLVTVSS
rHC44
640
EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYGVSWVRQAPGK
GLEWVSMIWADGSTHYADTLKSRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWQHGPVAYWGQGTLVTVSS
rHC1_G9
641
EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYGVTWVRQAPGK
GLEWVSIIGADGATDYADSVKSRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWQHGPLGYWGQGTLVTVSS
H1 + H2_A6
642
EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYGVNWVRQAPGK
GLEWVSGITGDGITAYASTLKSRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWHHGPVAYWGQGTLVTVSS
H1 + H2_G2
643
EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYGVSWVRQAPGK
GLEWVSMISGDGFTDYASSVKSRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWHHGPVAYWGQGTLVTVSS
H1 + H2_G7
644
EVQLVESGGGLVQPGGSLRLSCAASGFTFSDYGVNWVRQAPGK
GLEWVSNIWGDGSTDYASSLKGRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWHHGPVAYWGQGTLVTVSS
H1 + H2_E10
645
EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYGVNWVRQAPGK
GLEWVSMIRADGSTDYADSLKGRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWHHGPVAYWGQGTLVTVSS
rHC1_E2
646
EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYGVTWVRQAPGK
GLEWVSMIRGDGSTDYADSVKGRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWHHGPVAYWGQGTLVTVSS
H1 + H2_A4
647
EVQLVESGGGLVQPGGSLRLSCAASGFTFSAYGVSWVRQAPGK
GLEWVSMIWRDGSTDYADSLKGRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWHHGPVAYWGQGTLVTVSS
H1 + H2_H3
648
EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYGVHWVRQAPGK
GLEWVSMIWGDGSTHYASSVKSRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWHHGPVAYWGQGTLVTVSS
rHC1_G1
649
EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYGVHWVRQAPGK
GLEWVSGISADGSTDYASSVKSRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWHHGPVAYWGQGTLVTVSS
H1 + H2_E8
650
EVQLVESGGGLVQPGGSLRLSCAASGFTFSHYGVNWVRQAPGK
GLEWVSMIGGDGFTDYASSVKGRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWHHGPVAYWGQGTLVTVSS
H1 + H2_C9
651
EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYGVNWVRQAPGK
GLEWVSMIRADGSTDYASSLKSRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWHHGPVAYWGQGTLVTVSS
rHC1_F7
652
EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYGVQWVRQAPGK
GLEWVSVISADGFTDYASSVKGRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWHHGPVAYWGQGTLVTVSS
rHC1_F6
653
EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYGVTWVRQAPGK
GLEWVSMIGADGSTDYASSVKSRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWQHGPVGYWGQGTLVTVSS
rHC22
654
EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYGVTWVRQAPGK
GLEWVSMIWADGSTDYADTVKGRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWQHGPVAYWGQGTLVTVSS
rHC2_G5
655
EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYGVSWVRQAPGK
GLEWVSLIRGDGYTDYASSVKGRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWHHGPVAYWGQGTLVTVSS
rHC1_C12
656
EVQLVESGGGLVQPGGSLRLSCAASGFTFSHYGVSWVRQAPGK
GLEWVSVIRADGVTDYASSVKGRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWQHGPVGYWGQGTLVTVSS
rHC3
657
EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYGVSWVRQAPGK
GLEWVSMIWADGSTHYASSLKGRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWQHGPVAYWGQGTLVTVSS
rHC1_F1
658
EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYGVQWVRQAPGK
GLEWVSRINGDGSTDYASTLKGRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWHHGPVAYWGQGTLVTVSS
H1 + H2_E11
659
EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYGVNWVRQAPGK
GLEWVSMIRSDGFTDYASSVKGRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWHHGPVAYWGQGTLVTVSS
H1 + H2_B8
660
EVQLVESGGGLVQPGGSLRLSCAASGFTFSNFGVNWVRQAPGK
GLEWVSMIWVDGSTDYADSVKSRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWHHGPVAYWGQGTLVTVSS
H1 + H2_G1
661
EVQLVESGGGLVQPGGSLRLSCAASGFTFSDYGVNWVRQAPGK
GLEWVSMIWGDGSTYYASSVKSRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWHHGPVAYWGQGTLVTVSS
H1 + H2_B3
662
EVQLVESGGGLVQPGGSLRLSCAASGFTFSHYGVSWVRQAPGK
GLEWVSMIRSDGFTDYASTVKGRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWHHGPVAYWGQGTLVTVSS
rHC1_D2
663
EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYGVTWVRQAPGK
GLEWVSMITGDGYTDYADTVKGRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWQHGPLAYWGQGTLVTVSS
rHC1_E12
664
EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYGVSWVRQAPGK
GLEWVSIIRADGLTDYADSVKSRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWHHGPVAYWGQGTLVTVSS
rHC1_B5
665
EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYGVSWVRQAPGK
GLEWVSLIRSDGSTDYASSVKSRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWHHGPVAYWGQGTLVTVSS
rHC1_D11
666
EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYGVTWVRQAPGK
GLEWVSMIRADGSTDYADSVKGRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWQHGPLAYWGQGTLVTVSS
H1 + H2_A7
667
EVQLVESGGGLVQPGGSLRLSCAASGFTFSDYGVIWVRQAPGK
GLEWVSMIGGDGSTYYDSSLKGRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWHHGPVAYWGQGTLVTVSS
rHC1_G3
668
EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYGVNWVRQAPGK
GLEWVSMIGSDGSTDYASSVKGRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWHHGPVAYWGQGTLVTVSS
H1 + H2_D5
669
EVQLVESGGGLVQPGGSLRLSCAASGFTFSYYGVHWVRQAPGK
GLEWVSGISGEGSTDYASSVKGRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWHHGPVAYWGQGTLVTVSS
H1 + H2_D1
670
EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYGVTWVRQAPGK
GLEWVSMIRGDGSTYYASSVKSRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWHHGPVAYWVKGTLVTVSS
rHC1_E7
671
EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYGVSWVRQAPGK
GLEWVSIIRGDGSTDYASSLKSRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWHHGPVAYWGQGTLVTVSS
rHC1_E11
672
EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYGVTWVRQAPGK
GLEWVSMIRADGTTDYASSVKGRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWHHGPVAYWGQGTLVTVSS
S4-55
673
EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYGVTWVRQAPGK
GLEWVSMIWADGSTDYASTVKGRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWQHGPVGYWGQGTLVTVSS
H1 + H2_C10
674
EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYGVSWVRQAPGK
GLEWVSMIRGDGSTYYADTLKGRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWHHGPVAYWGQGTLVTVSS
H1 + H2_G10
675
EVQLVESGGGLVQPGGSLRLSCAASGFTFSHFGVNWVRQAPGK
GLEWVSMIWADGSTSYADSVKSRFTISRDNSKNTLYLQMNSLR
AEDTAVYYCAREWHHGPVAYWGQGTLVTVSS
Table 20 provides a list of amino acid sequences of VL regions of affinity matured fully human TNF antibodies derived from hMAK195 Amino acid residues of individual CDRs of each VH sequence are indicated in bold.
TABLE 20
List of amino acid sequences of affinity
matured hMAK195 VL variants
SEQ
ID
Clone
NO:
VL
S3_92
676
DIQMTQSPSSLSASVGDRVTITCRASQKVSSAVAWYQQK
PGKAPKLLIYWASTRHTGVPSRFSGSGSGTDFTLTISSL
QPEDFATYYCQQHYHTPYTFGQGTKLEIK
S3_79
677
DIQMTQSPSSLSASVGDRVTITCKASQAVSTEVAWYQQK
PGKAPKLLIYCASTRQTGVPSRFSGSGSGTDFTLTISSL
QPEDFATYYCQQSYSAPYTFGQGTKLEIK
S3_68
678
DIQMTQSPSSLSASVGDRVTITCRASQVVSSAVAWYQQK
PGKAPKLLIYWASKRHTGVPSRFSGSGSGTDFTLTISSL
QPEDFATYYCQQHYSTPFTFGQGTKLEIK
S3_60
679
DIQMTQSPSSLSASVGDRVTITCRASQAVSSAVAWYQQK
PGKAPKLLIYWASTRHTGVPSRFSGSGSGTDFTLTISSL
QPEDFATYYCQQHYSTPFTFGQGTKLEIK
S4-63
680
DIQMTQSPSSLSASVGDRVTITCKASQKVSSALAWYQQK
PGKAPKLLIYWASALHSGVPSRFSGSGSGTDFTLTISSL
QPEDFATYYCQQHYRPPFTFGQGTKLEIK
S3_5
681
DIQMTQSPSSLSASVGDRVTITCRASQGVSSAVAWYQQK
PGKAPKLLIYWASTRHTGVPSRFSGSGSGTDFTLTISSL
QPEDFATYYCQQHYTTPFTFGQGTKLEIK
S3_44
682
DIQMTQSPSSLSASVGDRVTITCRASQGVSRALAWYQQK
PGKAPKLLIYWASTLHSGVPSRFSGSGSGTDFTLTISSL
QPEDFATYYCQQHYRAPFTFGQGTKLEIK
S3_53
683
DIQMTQSPSSLSASVGDRVTITCRASQAVSSAVAWYQQK
PGKAPKLLIYWASTRHTGVPSRFSGSGSGTDFTLTISSL
QPEDFATYYCQQHYHTPFTFGQGTKLEIK
S3_91
684
DIQMTQSPSSLSASVGDRVTITCKASQGVSSALAWYQQK
PGKAPKLLIYWASTRHSGVPSRFSGSGSGTDFTLTISSL
QPEDFATYYCQQHYRTPFTFGQGTKLEIK
S3_59
685
DIQMTQSPSSLSASVGDRVTITCKASQGVSSALAWYQQK
PGKAPKLLIYWASTRHSGVPSRFSGSGSGTDFTLTISSL
QPEDFATYYCQQHYSTPYTFGQGTKLEIK
S3_47
686
DIQMTQSPSSLSASVGDRVTITCKASQWVSSAVAWYQQK
PGKAPKLLIYWASTRQSGVPSRFSGSGSGTDFTLTISSL
QPEDFATYYCQQHYRIPFTFGQGTKLEIK
S3_70
687
DIQMTQSPSSLSASVGDRVTITCKASQAVSSALAWYQQK
PGKAPKLLIYWASTRHSGVPSRFSGSGSGTDFTLTISSL
QPEDFATYYCQQHYSTPYTFGQGTKLEIK
S3_56
688
DIQMTQSPSSLSASVGDRVTITCKASQRVSSAVAWYQQK
PGKAPKLLIYWASTLHTGVPSRFSGSGSGTDFTLTISSL
QPEDFATYYCQQHYSTPYTFGQGTKLEIK
S3_37
689
DIQMTQSPSSLSASVGDRVTITCKASQGVSSAVAWYQQK
PGKAPKLLIYWASTRHTGVPSRFSGSGSGTDFTLTISSL
QPEDFATYYCQQHYNTPFTFGQGTKLEIK
S3_36
690
DIQMTQSPSSLSASVGDRVTITCKASQKVSSAVAWYQQK
PGKAPKLLIYWASARHTGVPSRFSGSGSGTDFTLTISSL
QPEDFATYYCQQHYSTPFTFGQGTKLEIK
S3_67
691
DIQMTQSPSSLSASVGDRVTITCKASQTVXRAVAWYQQK
PGKAPKLLIYWASTRQSGVPSRFSGSGSGTDFTLTISSL
QPEDFATYYCQQSYSTPFTFGQGTKLEIK
S3_40
692
DIQMTQSPSSLSASVGDRVTITCRASQRVSSAVAWSQQK
PGKAPKLLIYWASTRHSGVPSRFSGSGSGTDFTLTISSL
QPEDFATYYCQQHYTTPYTFGQGTKLEIK
S3_73
693
DIQMTQSPSSLSASVGDRVTITCKASQAVSSAVAWYQQK
PGKAPKLLIYWASTRHSGVPSRFSGSGSGTDFTLTISSL
QPEDFATYYCQQHYSTPFTFGQGTKLEIK
S4-50
694
DIQMTQSPSSLSASVGDRVTITCKASQLVSSAVAWYQQK
PGKAPKLLIYWASALHTGVPSRFSGSGSGTDFTLTISSL
QPEDFATYYCQQHYSSPYTFGQGTKLEIK
S4-6
695
DIQMTQSPSSLSASVGDRVTITCKASQLVSSAVAWYQQK
PGKAPKLLIYWASTRHTGVPSRFSGSGSGTDFTLTISSL
QPEDFATYYCQQHYSTPFTFGQGTKLEIK
S3_19
696
DIQMTQSPSSLSASVGDRVTITCKASQKVSSAVAWYQQK
PGKAPKLLIYWASARHSGVPSRFSGSGSGTDFTLTISSL
QPEDFATYYCQQHYRSPFTFGQGTKLEIK
S3_83
697
DIQMTQSPSSLSASVGDRVTITCRASQAVSTALAWYQQK
PGKAPKLLIYSASTLQSGVPSRFSGSGSGTDFTLTISSL
QPEDFATYYCQQHYRSPFTFGQGTKLEIK
S3_78
698
DIQMTQSPSSLSASVGDRVTITCKASQYVGGAVAWYQQK
PGKAPKLLIYQASTLQTGVPSRFSGSGSGTDFTLTISSL
QPEDFATYYCQQHISKPFTFGQGTKLEIK
S4-19
699
DIQMTQSPSSLSASVGDRVTITCKASQLVSSAVAWYQQK
PGKAPKLLIYWASTLHTGVPSRFSGSGSGTDFTLTISSL
QPEDFATYYCQQHYRTPFTFGQGTKLEIK
S3_58
700
DIQMTQSPSSLSASVGDRVTITCKASQSVNGALAWYQQK
PGKAPKLLIYRASTRQTGVPSRFSGSGSGTDFTLTISSL
QPEDFATYYCQQHYSIPFTFGQGTKLEIK
S4-31
701
DIQMTQSPSSLSASVGDRVTITCRASQGVSSALAWYQQK
PGKAPKLLIYWASALHSGVPSRFSGSGSGTDFTLTISSL
QPEDFATYYCQQHYSAPFTFGQGTKLEIK
S3_31
702
DIQMTQSPSSLSASVGDRVTITCKASQAVSSSVAWYQQK
PGKAPKLLIYGASTLQSGVPSRFSGSGSGTDFTLTISSL
QPEDFATYYCQQHYNEPYTFGQGTKLEIK
S3_13
703
DIQMTQSPSSLSASVGDRVTITCKASQKVSSAVAWYQQK
PGKAPKLLIYWASARHSGVPSRFSGSGSGTDFTLTISSL
QPEDFATYYCQQHYRTPYTFGQGTKLEIK
S4-40
704
DIQMTQSPSSLSASVGDRVTITCRASQLVSSAVAWYQQK
PGKAPKLLIYWASTRHSGVPSRFSGSGSGTDFTLTISSL
QPEDFATYYCQQHYRTPFSFGQGTKLEIK
S3_26
705
DIQMTQSPSSLSASVGDRVTITCRASQAVSSAVAWYQQK
PGKAPKLLIYWASKRQTGVPSRFSGSGSGTDFTLTISSL
QPEDFATYYCQQHYISPYTFGQGTKLEIK
S3_33
706
DIQMTQSPSSLSASVGDRVTITCKASQGVRSALAWYQQK
PGKAPKLLIYWASTRHTGVPSRFSGSGSGTDFTLTISSL
QPEDFATYYCQQSYSAPYTFGQGTKLEIK
S3_28
707
DIQMTQSPSSLSASVGDRVTITCKASQTVSNAVAWYQQK
PGKAPKLLIYWASTRHSGVPSRFSGSGSGTDFTLTISSL
QPEDFATYYCQQHYSTPFTFGQGTKLEIK
S4-74
708
DIQMTQSPSSLSASVGDRVTITCRASQLVSSAVAWYQQK
PGKAPKLLIYWASARHTGVPSRFSGSGSGTDFTLTISSL
QPEDFATYYCQQHYRTPFTFGQGTKLEIK
S3_84
709
DIQMTQSPSSLSASVGDRVTITCKASQPVRSAVAWYQQK
PGKAPKLLIYSASTRQTGVPSRFSGSGSGTDFTLTISSL
QPEDFATYYCQQSYSIPFTFGQGTKLEIK
S4-54
710
DIQMTQSPSSLSASVGDRVTITCRASQLVSSAVAWYQQK
PGKAPKLLIYWASARHTGVPSRFSGSGSGTDFTLTISSL
QPEDFATYYCQQHYKTPFSFGQGTKLEIK
S3_23
711
DIQMTQSPSSLSASVGDRVTITCRASQAVSSAVAWYQQK
PGKAPKLLIYWASSRHTGVPSRFSGSGSGTDFTLTISSL
QPEDFATYYCQQHYSTPFTFGQGTKLEIK
S3_55
712
DIQMTQSPSSLSASVGDRVTITCKASQTVGRAVAWYQQK
PGKAPKLLIYWASTRQSGVPSRFSGSGSGTDFTLTISSL
QPEDFATYYCQQSYSTPFTFGQGTKLEIK
S4-34
713
DIQMTQSPSSLSASVGDRVTITCRASQLVSSAVAWYQQK
PGKAPKLLIYWASTRHTGVPSRFSGSGSGTDFTLTISSL
QPEDFATYYCQQHYRTPFTFGQGTKLEIK
S3_76
714
DIQMTQSPSSLSASVGDRVTITCRASQKVSNAVAWYQQK
PGKAPKLLIYWASTRHTGVPSRFSGSGSGTDFTLTISSL
QPEDFATYYCQQHYNSPFTFGQGTKLEIK
S4-12
715
DIQMTQSPSSLSASVGDRVTITCRASQLVSSAVAWYQQK
PGKAPKLLIYWASARHTGVPSRFSGSGSGTDFTLTISSL
QPEDFATYYCQQHYKTPFTFGQGTKLEIK
S3_86
716
DIQMTQSPSSLSASVGDRVTITCRASQRVSSAVAWYQQK
PGKAPKLLIYWASTRHSGVPSRFSGSGSGTDFTLTISSL
QPEDFATYYCQQHYTTPYTFGQGTKLEIK
S3_61
717
DIQMTQSPSSLSASVGDRVTITCKASQRVSSAVAWYQQK
PGKAPKLLIYWASNRHSGVPSRFSGSGSGTDFTLTISSL
QPEDFATYYCQQHYSTPFTFGQGTKLEIK
S3_18
718
DIQMTQSPSSLSASVGDRVTITCKASQLVSSALAWYQQK
PGKAPKLLIYWASTRQSGVPSRFSGSGSGTDFTLTISSL
QPEDFATYYCQQHYRTPFTFGQGTKLEIK
S3_72
719
DIQMTQSPSSLSASVGDRVTITCKASQLVSSALAWYQQK
PGKAPKLLIYWASTRHTGVPSRFSGSGSGTDFTLTISSL
QPEDFATYYCQQHYRNPFTFGQGTKLEIK
S3_41
720
DIQMTQSPSSLSASVGDRVTITCKASQAVSSALAWYQQK
PXKAPKLLIYWASSRQSGVPSRFSGSGSGTDFTLTISSL
QPEDFATYYCQQHYRTPFTFGQGTKLEIK
S4-24
721
DIQMTQSPSSLSASVGDRVTITCRASQLVSSAVAWYQQK
PGKAPKLLIYWASTLHTGVPSRFSGSGSGTDFTLTISSL
QPEDFATYYCQQHYRTPFTFGQGTKLEIK
S4-17
722
DIQMTQSPSSLSASVGDRVTITCRASQLVSSAVAWYQQK
PGKAPKLLIYWASTRHSGVPSRFSGSGSGTDFTLTISSL
QPEDFATYYCQQHYRTPFTFGQGTKLEIK
S3_90
723
DIQMTQSPSSLSASVGDRVTITCKASQPVSGAVAWYQQK
PGKAPKLLIYWASTRHTGVPSRFSGSGSGTDFTLTISSL
QPEDFATYYCQQHYRASYTFGQGTKLEIK
S3_87
724
DIQMTQSPSSLSASVGDRVTITCRASQKVSSAVAWYQQK
PGKAPKLLIYWASARHTGVPSRFSGSGSGTDFTLTISSL
QPEDFATYYCQQHYRTPYTFGQGTKLEIK
S3_66
725
DIQMTQSPSSLSASVGDRVTITCRASQRVSSAVAWYQQK
PGKAPKLLIYWASTRHSGVPSRFSGSGSGTDFTLTISSL
QPEDFATYYCQQHYTTPYTFGQGTKLEIK
S4-18
726
DIQMTQSPSSLSASVGDRVTITCRASQLVSSAVAWYQQK
PGKAPKLLIYWASTLHSGVPSRFSGSGSGTDFTLTISSL
QPEDFATYYCQQHYSTPFTFGQGTKLEIK
S3_4
727
DIQMTQSPSSLSASVGDRVTITCRASQAVSSAVAWYQQK
PGKAPKLLIYWASARHTGVPSRFSGSGSGTDFTLTISSL
QPEDFATYYCQQHYSSPYTFGQGTKLEIK
S3_64
728
DIQMTQSPSSLSASVGDRVTITCKASQPVSSAVAWYQQK
PGKAPKLLIYWASTLHSGVPSRFSGSGSGTDFTLTISSL
QPEDFATYYCQQHYRTPFTFGQGTKLEIK
S3_62
729
DIQMTQSPSSLSASVGDRVTITCRASQLVSSAVAWYQQK
PGKAPKLLIYWASTRHTGVPSRFSGSGSGTDFTLTISSL
QPEDFATYYCQQHYRTPYTFGQGTNLEIK
S3_29
730
DIQMTQSPSSLSASVGDIVTITCKASQLVSSAVAWYQQK
PGKAPKLLIYWASTRHTGVPSRFSGSGSGTDFTLTISSL
QPEDFATYYCQQHYRTPYTFGQGTKLEIK
S3_65
731
DIQMTQSPSSLSASVGDRVTITCKASQLVSSAVAWYQQK
PGKAPKLLIYWASMRHSGVPSRFSGSGSGTDFTLTISSL
QPEDFATYYCQQHYSSPFTFGQGTKLEIK
S3_81
732
DIQMTQSPSSLSASVGDRVTITCKASQTVSSAVAWYQQK
PGKAPKLLIYWASTRHSGVPSRFSGSGSGTDFTLTISSL
QPEDFATYYCQQHYRAPYTFGQGTKLEIK
S3_39
733
DIQMTQSPSSLSASVGDRVTITCKASQRVSSALAWYQQK
PGKAPKLLIYWASTRHTGVPSRFSGSGSGTDFTLTISSL
QPEDFATYYCQQHYSTPFTFGQGTKLEIK
S3_49
734
DIQMTQSPSSLSASVGDRVTITCRASQLVSNAVAWYQQK
PGKAPKLLIYWASTRHTGVPSRFSGSGSGTDFTLTISSL
QPEDFATYYCQQHYSSPFTFGQGTKLEIK
S3_85
735
DIQMTQSPSSLSASVGDRVTITCRASQLVSSAVAWYQQK
PGKAPKLLIYWASARHSGVPSRFSGSGSGTDFTLTISSL
QPEDFATYYCQQHYSTPFTFGQGTKLEIK
S3_82
736
DIQMTQSPSSLSASVGDRVTITCKASQLVSSAVAWYQQK
PGKAPKLLIYWASTRHSGVPSRFSGSGSGTDFTLTISSL
QPEDFATYYCQQHYTTPFTFGQGTKLEIK
S3_93
737
DIQMTQSPSSLSASVGDRVTITCKASQRVSSAVAWYQQK
PGKAPKLLIYWASTRHTGVPSRFSGSGSGTDFTLTISSL
QPEDFATYYCQQHYSTPFTFGQGTKLEIK
TABLE 21
Amino acid residues observed in affinity matured hMAK-195.
hMAK195 Heavy chain variable region (SEQ ID NO: 1075)
hMAK195VH
EVQLVESGGGLVQPGGSLRLSCAASGFTFSDYGVNWVRQAPGKGLEWVSMIWGDGSTD
NFS T I RAG T A
HLN S V GSE F H
YS H L SDA A V
IR Q R AEV Y S
Y K LVG W N
S NY G
YDSTLKSRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAREWHHGPVAYWGQGTLVTVSS
ADSV G HSQQRTLDS
QLRPASGVF
LCLLVQDGC
YRYNWAETN
DFPYEKW P
NDARS R I
TYVTP P H
PPDDI A
AICA I
SG C
R
hMAK195 Light chain variable region (SEQ ID NO: 1076)
hMAK195VL
DIQMTQSPSSLSASVGDRVTITCKASQAVSSAVAWYQQKPGKAPKLLIYWASTRHTG
R S RRPL S SLQS
V TNT R I T
G IGG L L A
D NCV C K E
T CTS Q A F
P KIR G R
VPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQHYSTPFTFGQGTKLEIK
SNRSTY
FGPR
DTML
GIIQ
HCAA
S
The tables below provide a list of humanized MAK-195 antibodies that were converted into IgG proteins for characterization.
TABLE 22
VH sequences of IgG converted clones
Protein
region
SEQ ID NO:
Sequence
123456789012345678901234567890
A8
738
EVQLVESGGGLVQPGGSLRLSCAASGFTFS
VH
NYGVNWVRQAPGKGLEWVSMIAADGFTDYA
SSVKGRFTISRDNSKNTLYLQMNSLRAEDT
AVYYCAREWHHGPVAYWGQGTLVTVSS
A8
CDR-H1
Residues 31-35
NYGVN
VH
of SEQ ID
NO.: 738
A8
CDR-H2
Residues 50-65
MIAADGFTDYASSVKG
VH
of SEQ ID
NO.: 738
A8
CDR-H3
Residues 98-106
EWHHGPVAY
VH
of SEQ ID
NO.: 738
B5
739
EVQLVESGGGLVQPGGSLRLSCAASGFTFS
VH
NYGVSWVRQAPGKGLEWVSLIRGDGSTDYA
SSLKGRFTISRDNSKNTLYLQMNSLRAEDT
AVYYCAREWHHGPVAYWGQGTLVTVSS
B5
CDR-H1
Residues 31-35
NYGVS
VH
of SEQ ID
NO.: 739
B5
CDR-H2
Residues 50-65
LIRGDGSTDYASSLKG
VH
of SEQ ID
NO.: 739
B5
CDR-H3
Residues 98-106
EWHHGPVAY
VH
of SEQ ID
NO.: 739
rHC44
740
EVQLVESGGGLVQPGGSLRLSCAASGFTFS
VH
NYGVSWVRQAPGKGLEWVSMIWADGSTHYA
DTLKSRFTISRDNSKNTLYLQMNSLRAEDT
AVYYCAREWQHGPVAYWGQGTLVTVSS
rHC44
CDR-H1
Residues 31-35
NYGVS
VH
of SEQ ID
NO.: 740
rHC44
CDR-H2
Residues 50-65
MIWADGSTHYADTLKS
VH
of SEQ ID
NO.: 740
rHC44
CDR-H3
Residues 98-106
EWQHGPVAY
VH
of SEQ ID
NO.: 740
rHC22
741
EVQLVESGGGLVQPGGSLRLSCAASGFTFS
VH
NYGVTWVRQAPGKGLEWVSMIWADGSTDYA
DTVKGRFTISRDNSKNTLYLQMNSLRAEDT
AVYYCAREWQHGPVAYWGQGTLVTVSS
rHC22
CDR-H1
Residues 31-35
NYGVT
VH
of SEQ ID
NO.: 741
rHC22
CDR-H2
Residues 50-65
MIWADGSTDYADTVKG
VH
of SEQ ID
NO.: 741
rHC22
CDR-H3
Residues 98-106
EWQHGPVAY
VH
of SEQ ID
NO.: 741
rHC81
742
EVQLVESGGGLVQPGGSLRLSCAASGFTFS
VH
NYGVTWVRQAPGKGLEWVSMIWADGSTHYA
DSVKSRFTISRDNSKNTLYLQMNSLRAEDT
AVYYCAREWQHGPLAYWGQGTLVTVSS
rHC81
CDR-H1
Residues 31-35
NYGVT
VH
of SEQ ID
NO.: 742
rHC81
CDR-H2
Residues 50-65
MIWADGSTHYADSVKS
VH
of SEQ ID
NO.: 742
rHC81
CDR-H3
Residues 98-106
EWQHGPLAY
VH
of SEQ ID
NO.: 742
rHC18
743
EVQLVESGGGLVQPGGSLRLSCAASGFTFS
VH
NYGVTWVRQAPGKGLEWVSMIWSDGSTDYA
SSVKGRFTISRDNSKNTLYLQMNSLRAEDT
AVYYCAREWQHGPVAYWGQGTLVTVSS
rHC18
CDR-H1
Residues 31-35
NYGVT
VH
of SEQ ID
NO.: 743
rHC18
CDR-H2
Residues 50-65
MIWSDGSTDYASSVKG
VH
of SEQ ID
NO.: 743
rHC18
CDR-H3
Residues 98-106
EWQHGPVAY
VH
of SEQ ID
NO.: 743
rHC14
744
EVQLVESGGGLVQPGGSLRLSCAASGFTFS
VH
NYGVTWVRQAPGKGLEWVSMIWADGSTHYA
SSLKGRFTISRDNSKNTLYLQMNSLRAEDT
AVYYCAREWQHGPAAYWGQGTLVTVSS
rHC14
CDR-H1
Residues 31-35
NYGVT
VH
of SEQ ID
NO.: 744
rHC14
CDR-H2
Residues 50-65
MIWADGSTHYASSLKG
VH
of SEQ ID
NO.: 744
rHC14
CDR-H3
Residues 98-106
EWQHGPAAY
VH
of SEQ ID
NO.: 744
rHC3
745
EVQLVESGGGLVQPGGSLRLSCAASGFTFS
VH
NYGVSWVRQAPGKGLEWVSMIWADGSTHYA
SSLKGRFTISRDNSKNTLYLQMNSLRAEDT
AVYYCAREWQHGPVAYWGQGTLVTVSS
rHC3
CDR-H1
Residues 31-35
NYGVS
VH
of SEQ ID
NO.: 745
rHC3
CDR-H2
Residues 50-65
MIWADGSTHYASSLKG
VH
of SEQ ID
NO.: 745
rHC3
CDR-H3
Residues 98-106
EWQHGPVAY
VH
of SEQ ID
NO.: 745
rHC19
746
EVQLVESGGGLVQPGGSLRLSCAASGFTFS
VH
NYGVTWVRQAPGKGLEWVSMIWADGSTHYA
SSVKGRFTISRDNSKNTLYLQMNSLRAEDT
AVYYCAREWQHGPAAYWGQGTLVTVSS
rHC19
CDR-H1
Residues 31-35
NYGVT
VH
of SEQ ID
NO.: 746
rHC19
CDR-H2
Residues 50-65
MIWADGSTHYASSVKG
VH
of SEQ ID
NO.: 746
rHC19
CDR-H3
Residues 98-106
EWQHGPAAY
VH
of SEQ ID
NO.: 746
rHC34
747
EVQLVESGGGLVQPGGSLRLSCAASGFTFS
VH
NYGVTWVRQAPGKGLEWVSMIWADGSTHYA
SSVKGRFTISRDNSKNTLYLQMNSLRAEDT
AVYYCAREWQHGPVAYWGQGTLVTVSS
rHC34
CDR-H1
Residues 31-35
NYGVT
VH
of SEQ ID
NO.: 747
rHC34
CDR-H2
Residues 50-65
MIWADGSTHYASSVKG
VH
of SEQ ID
NO.: 747
rHC34
CDR-H3
Residues 98-106
EWQHGPSAY
VH
of SEQ ID
NO.: 747
rHC83
748
EVQLVESGGGLVQPGGSLRLSCAASGFTFS
VH
NYGVTWVRQAPGKGLEWVSMIWADGSTHYA
SSVKGRFTISRDNSKNTLYLQMNSLRAEDT
AVYYCAREWQHGPVAYWGQGTLVTVSS
rHC83
CDR-H1
Residues 31-35
NYGVT
VH
of SEQ ID
NO.: 748
rHC83
CDR-H2
Residues 50-65
MIWADGSTHYASSVKG
VH
of SEQ ID
NO.: 748
rHC83
CDR-H3
Residues 98-106
EWQHGPVAY
VH
of SEQ ID
NO.: 748
S4-19
749
EVQLVESGGGLVQPGGSLRLSCAASGFTFS
VH
NYGVEWVRQAPGKGLEWVSGIWADGSTHYA
DTVKSRFTISRDNSKNTLYLQMNSLRAEDT
AVYYCAREWQHGPVAYWGQGTLVTVSS
S4-19
CDR-H1
Residues 31-35
NYGVE
VH
of SEQ ID
NO.: 749
S4-19
CDR-H2
Residues 50-65
GIWADGSTHYADTVKS
VH
of SEQ ID
NO.: 749
S4-19
CDR-H3
Residues 98-106
EWQHGPVAY
VH
of SEQ ID
NO.: 749
S4-50
750
EVQLVESGGGLVQPGGSLRLSCAASGFTFS
VH
NYGVEWVRQAPGKGLEWVSGIWADGSTHYA
DTVKSRFTISRDNSKNTLYLQMNSLRAEDT
AVYYCAREWQHGPVGYWGQGTLVTVSS
S4-50
CDR-H1
Residues 31-35
NYGVE
VH
of SEQ ID
NO.: 750
S4-50
CDR-H2
Residues 50-65
GIWADGSTHYADTVKS
VH
of SEQ ID
NO.: 750
S4-50
CDR-H3
Residues 98-106
EWQHGPVGY
VH
of SEQ ID
NO.: 750
S4-63
751
EVQLVESGGGLVQPGGSLRLSCAASGFTFS
VH
NYGVEWVRQAPGKGLEWVSGIWADGSTHYA
DTVKSRFTISRDNSKNTLYLQMNSLRAEDT
AVYYCAREWQHGPVGYWGQGTLVTVSS
S4-63
CDR-H1
Residues 31-35
NYGVE
VH
of SEQ ID
NO.: 751
S4-63
CDR-H2
Residues 50-65
GIWADGSTHYADTVKS
VH
of SEQ ID
NO.: 751
S4-63
CDR-H3
Residues 98-106
EWQHGPVGY
VH
of SEQ ID
NO.: 751
S4-55
752
EVQLVESGGGLVQPGGSLRLSCAASGFTFS
VH
NYGVTWVRQAPGKGLEWVSMIWADGSTDYA
STVKGRFTISRDNSKNTLYLQMNSLRAEDT
AVYYCAREWQHGPVGYWGQGTLVTVSS
S4-55
CDR-H1
Residues 31-35
NYGVT
VH
of SEQ ID
NO.: 752
S4-55
CDR-H2
Residues 50-65
MIWADGSTDYASTVKG
VH
of SEQ ID
NO.: 752
S4-55
CDR-H3
Residues 98-106
EWQHGPVGY
VH
of SEQ ID
NO.: 752
S4-6
753
EVQLVESGGGLVQPGGSLRLSCAASGFTFS
VH
NYGVTWVRQAPGKGLEWVSMIWADGSTHYA
SSVKGRFTISRDNSKNTLYLQMNSLRAEDT
AVYYCAREWQHGPVAYWGQGTLVTVSS
S4-6
CDR-H1
Residues 31-35
NYGVT
VH
of SEQ ID
NO.: 753
S4-6
CDR-H2
Residues 50-65
MIWADGSTHYASSVKG
VH
of SEQ ID
NO.: 753
S4-6
CDR-H3
Residues 98-106
EWQHGPVAY
VH
of SEQ ID
NO.: 753
S4-18
754
EVQLVESGGGLVQPGGSLRLSCAASGFTFS
VH
NYGVTWVRQAPGKGLEWVSMIWADGSTHYA
DSVKSRFTISRDNSKNTLYLQMNSLRAEDT
AVYYCAREWQHGPLAYWGQGTLVTVSS
S4-18
CDR-H1
Residues 31-35
NYGVT
VH
of SEQ ID
NO.: 754
S4-18
CDR-H2
Residues 50-65
MIWADGSTHYADSVKS
VH
of SEQ ID
NO.: 754
S4-18
CDR-H3
Residues 98-106
EWQHGPLAY
VH
of SEQ ID
NO.: 754
S4-31
755
EVQLVESGGGLVQPGGSLRLSCAASGFTFS
VH
NYGVQWVRQAPGKGLEWVSGIGADGSTAYA
SSLKGRFTISRDNSKNTLYLQMNSLRAEDT
AVYYCAREWQHSGLAYWGQGTLVTVSS
S4-31
CDR-H1
Residues 31-35
NYGVQ
VH
of SEQ ID
NO.: 755
S4-31
CDR-H2
Residues 50-65
GIGADGSTAYASSLKG
VH
of SEQ ID
NO.: 755
S4-31
CDR-H3
Residues 98-106
EWQHSGLAY
VH
of SEQ ID
NO.: 755
S4-34
756
EVQLVESGGGLVQPGGSLRLSCAASGFTFS
VH
NYGVSWVRQAPGKGLEWVSMIWADGSTHYA
DTVKGRFTISRDNSKNTLYLQMNSLRAEDT
AVYYCAREWQHGPLAYWGQGTLVTVSS
S4-34
CDR-H1
Residues 31-35
NYGVS
VH
of SEQ ID
NO.: 756
S4-34
CDR-H2
Residues 50-65
MIWADGSTHYADTVKG
VH
of SEQ ID
NO.: 756
S4-34
CDR-H3
Residues 98-106
EWQHGPLAY
VH
of SEQ ID
NO.: 756
S4-74
757
EVQLVESGGGLVQPGGSLRLSCAASGFTFS
VH
NYGVTWVRQAPGKGLEWVSMIWADGSTHYA
DTVKGRFTISRDNSKNTLYLQMNSLRAEDT
AVYYCAREWQHGPLAYWGQGTLVTVSS
S4-74
CDR-H1
Residues 31-35
NYGVT
VH
of SEQ ID
NO.: 757
S4-74
CDR-H2
Residues 50-65
MIWADGSTHYADTVKG
VH
of SEQ ID
NO.: 757
S4-74
CDR-H3
Residues 98-106
EWQHGPLAY
VH
of SEQ ID
NO.: 757
S4-12
758
EVQLVESGGGLVQPGGSLRLSCAASGFTFS
VH
NYGVTWVRQAPGKGLEWVSMIWADGSTHYA
SSVKGRFTISRDNSKNTLYLQMNSLRAEDT
AVYYCAREWQHGPVAYWGQGTLVTVSS
S4-12
CDR-H1
Residues 31-35
NYGVT
VH
of SEQ ID
NO.: 758
S4-12
CDR-H2
Residues 50-65
MIWADGSTHYASSVKG
VH
of SEQ ID
NO.: 758
S4-12
CDR-H3
Residues 98-106
EWQHGPVAY
VH
of SEQ ID
NO.: 758
S4-54
759
EVQLVESGGGLVQPGGSLRLSCAASGFTFS
VH
NYGVTWVRQAPGKGLEWVSMIWADGSTHYA
SSVKGRFTISRDNSKNTLYLQMNSLRAEDT
AVYYCAREWQHGPVAYWGQGTLVTVSS
S4-54
CDR-H1
Residues 31-35
NYGVT
VH
of SEQ ID
NO.: 759
S4-54
CDR-H2
Residues 50-65
MIWADGSTHYASSVKG
VH
of SEQ ID
NO.: 759
S4-54
CDR-H3
Residues 98-106
EWQHGPVAY
VH
of SEQ ID
NO.: 759
S4-17
760
EVQLVESGGGLVQPGGSLRLSCAASGFTFS
VH
NYGVTWVRQAPGKGLEWVSMIWADGSTHYA
SSVKGRFTISRDNSKNTLYLQMNSLRAEDT
AVYYCAREWQHGPVAYWGQGTLVTVSS
S4-17
CDR-H1
Residues 31-35
NYGVT
VH
of SEQ ID
NO.: 760
S4-17
CDR-H2
Residues 50-65
MIWADGSTHYASSVKG
VH
of SEQ ID
NO.: 760
S4-17
CDR-H3
Residues 98-106
EWQHGPVAY
VH
of SEQ ID
NO.: 760
S4-40
761
EVQLVESGGGLVQPGGSLRLSCAASGFTFS
VH
NYGVTWVRQAPGKGLEWVSMIWADGSTHYA
SSVKGRFTISRDNSKNTLYLQMNSLRAEDT
AVYYCAREWQHGPVAYWGQGTLVTVSS
S4-40
CDR-H1
Residues 31-35
NYGVT
VH
of SEQ ID
NO.: 761
S4-40
CDR-H2
Residues 50-65
MIWADGSTHYASSVKG
VH
of SEQ ID
NO.: 761
S4-40
CDR-H3
Residues 98-106
EWQHGPVAY
VH
of SEQ ID
NO.: 761
S4-24
762
EVQLVESGGGLVQPGGSLRLSCAASGFTFS
VH
NYGVTWVRQAPGKGLEWVSMIWADGSTHYA
SSVKGRFTISRDNSKNTLYLQMNSLRAEDT
AVYYCAREWQHGPVAYWGQGTLVTVSS
S4-24
CDR-H1
Residues 31-35
NYGVT
VH
of SEQ ID
NO.: 762
S4-24
CDR-H2
Residues 50-65
MIWADGSTHYASSVKG
VH
of SEQ ID
NO.: 762
S4-24
CDR-H3
Residues 98-106
EWQHGPVAY
VH
of SEQ ID
NO.: 762
TABLE 23
VL sequences of IgG converted clones
Protein
region
SEQ ID NO:
Sequence
123456789012345678901234567890
hMAK195
763
DIQMTQSPSSLSASVGDRVTITCKASQAVS
VL.1
SAVAWYQQKPGKAPKLLIYWASTRHTGVPS
VL
RFSGSGSGTDFTLTISSLQPEDFATYYCQQ
HYSTPFTFGQGTKLEIKR
hMAK195
CDR-L1
Residues 24-34
KASQAVSSAVA
VL.1
of SEQ ID
VL
NO.: 763
hMAK195
CDR-L2
Residues 50-56
WASTRHT
VL.1
of SEQ ID
VL
NO.: 763
hMAK195
CDR-L3
Residues 89-97
QQHYSTPFT
VL.1
of SEQ ID
VL
NO.: 763
S4-24
764
DIQMTQSPSSLSASVGDRVTITCRASQLVS
VL
SAVAWYQQKPGKAPKLLIYWASTLHTGVPS
RFSGSGSGTDFTLTISSLQPEDFATYYCQQ
HYRTPFTFGQGTKLEIKR
S4-24
CDR-L1
Residues 24-34
RASQLVSSAVA
VL
of SEQ ID
NO.: 764
S4-24
CDR-L2
Residues 50-56
WASTLHT
VL
of SEQ ID
NO.: 764
S4-24
CDR-L3
Residues 89-97
QQHYRTPFT
VL
of SEQ ID
NO.: 764
S4-40
765
DIQMTQSPSSLSASVGDRVTITCRASQLVS
VL
SAVAWYQQKPGKAPKLLIYWASTRHSGVPS
RFSGSGSGTDFTLTISSLQPEDFATYYCQQ
HYRTPFSFGQGTKLEIKR
S4-40
CDR-L1
Residues 24-34
RASQLVSSAVA
VL
of SEQ ID
NO.: 765
S4-40
CDR-L2
Residues 50-56
WASTRHS
VL
of SEQ ID
NO.: 765
S4-40
CDR-L3
Residues 89-97
QQHYRTPFS
VL
of SEQ ID
NO.: 765
S4-17
766
DIQMTQSPSSLSASVGDRVTITCRASQLVS
VL
SAVAWYQQKPGKAPKLLIYWASTRHSGVPS
RFSGSGSGTDFTLTISSLQPEDFATYYCQQ
HYRTPFTFGQGTKLEIKR
S4-17
CDR-L1
Residues 24-34
RASQLVSSAVA
VL
of SEQ ID
NO.: 766
S4-17
CDR-L2
Residues 50-56
WASTRHS
VL
of SEQ ID
NO.: 766
S4-17
CDR-L3
Residues 89-97
QQHYRTPFT
VL
of SEQ ID
NO.: 766
S4-54
767
DIQMTQSPSSLSASVGDRVTITCRASQLVS
VL
SAVAWYQQKPGKAPKLLIYWASARHTGVPS
RFSGSGSGTDFTLTISSLQPEDFATYYCQQ
HYKTPFSFGQGTKLEIKR
S4-54
CDR-L1
Residues 24-34
RASQLVSSAVA
VL
of SEQ ID
NO.: 767
S4-54
CDR-L2
Residues 50-56
WASARHT
VL
of SEQ ID
NO.: 767
S4-54
CDR-L3
Residues 89-97
QQHYKTPFS
VL
of SEQ ID
NO.: 767
S4-12
768
DIQMTQSPSSLSASVGDRVTITCRASQLVS
VL
SAVAWYQQKPGKAPKLLIYWASARHTGVPS
RFSGSGSGTDFTLTISSLQPEDFATYYCQQ
HYKTPFTFGQGTKLEIKR
S4-12
CDR-L1
Residues 24-34
RASQLVSSAVA
VL
of SEQ ID
NO.: 768
S4-12
CDR-L2
Residues 50-56
WASARHT
VL
of SEQ ID
NO.: 768
S4-12
CDR-L3
Residues 89-97
QQHYKTPFT
VL
of SEQ ID
NO.: 768
S4-74
769
DIQMTQSPSSLSASVGDRVTITCRASQLVS
VL
SAVAWYQQKPGKAPKLLIYWASARHTGVPS
RFSGSGSGTDFTLTISSLQPEDFATYYCQQ
HYRTPFTFGQGTKLEIKR
S4-74
CDR-L1
Residues 24-34
RASQLVSSAVA
VL
of SEQ ID
NO.: 769
S4-74
CDR-L2
Residues 50-56
WASARHT
VL
of SEQ ID
NO.: 769
S4-74
CDR-L3
Residues 89-97
QQHYRTPFT
VL
of SEQ ID
NO.: 769
S4-34
770
DIQMTQSPSSLSASVGDRVTITCRASQLVS
VL
SAVAWYQQKPGKAPKLLIYWASTRHTGVPS
RFSGSGSGTDFTLTISSLQPEDFATYYCQQ
HYRTPFTFGQGTKLEIKR
S4-34
CDR-L1
Residues 24-34
RASQLVSSAVA
VL
of SEQ ID
NO.: 770
S4-34
CDR-L2
Residues 50-56
WASTRHT
VL
of SEQ ID
NO.: 770
S4-34
CDR-L3
Residues 89-97
QQHYRTPFT
VL
of SEQ ID
NO.: 770
S4-31
771
DIQMTQSPSSLSASVGDRVTITCRASQGVS
VL
SALAWYQQKPGKAPKLLIYWASALHSGVPS
RFSGSGSGTDFTLTISSLQPEDFATYYCQQ
HYSAPFTFGQGTKLEIKR
S4-31
CDR-L1
Residues 24-34
RASQGVSSALA
VL
of SEQ ID
NO.: 771
S4-31
CDR-L2
Residues 50-56
WASALHS
VL
of SEQ ID
NO.: 771
S4-31
CDR-L3
Residues 89-97
QQHYSAPFT
VL
of SEQ ID
NO.: 771
S4-18
772
DIQMTQSPSSLSASVGDRVTITCRASQLVS
VL
SAVAWYQQKPGKAPKLLIYWASTLHSGVPS
RFSGSGSGTDFTLTISSLQPEDFATYYCQQ
HYSTPFTFGQGTKLEIKR
S4-18
CDR-L1
Residues 24-34
RASQLVSSAVA
VL
of SEQ ID
NO.: 772
S4-18
CDR-L2
Residues 50-56
WASTLHS
VL
of SEQ ID
NO.: 772
S4-18
CDR-L3
Residues 89-97
QQHYSTPFT
VL
of SEQ ID
NO.: 772
S4-6
773
DIQMTQSPSSLSASVGDRVTITCKASQLVS
VL
SAVAWYQQKPGKAPKLLIYWASTRHTGVPS
RFSGSGSGTDFTLTISSLQPEDFATYYCQQ
HYSTPFTFGQGTKLEIKR
S4-6
CDR-L1
Residues 24-34
KASQLVSSAVA
VL
of SEQ ID
NO.: 773
S4-6
CDR-L2
Residues 50-56
WASTRHT
VL
of SEQ ID
NO.: 773
S4-6
CDR-L3
Residues 89-97
QQHYSTPFT
VL
of SEQ ID
NO.: 773
S4-55
774
DIQMTQSPSSLSASVGDRVTITCKASQLVS
VL
SAVAWYQQKPGKAPKLLIYWASTLHTGVPS
RFSGSGSGTDFTLTISSLQPEDFATYYCQQ
HYRTPFTFGQGTKLEIKR
S4-55
CDR-L1
Residues 24-34
KASQLVSSAVA
VL
of SEQ ID
NO.: 774
S4-55
CDR-L2
Residues 50-56
WASTLHT
VL
of SEQ ID
NO.: 774
S4-55
CDR-L3
Residues 89-97
QQHYRTPFT
VL
of SEQ ID
NO.: 774
S4-63
775
DIQMTQSPSSLSASVGDRVTITCKASQKVS
VL
SALAWYQQKPGKAPKLLIYWASALHSGVPS
RFSGSGSGTDFTLTISSLQPEDFATYYCQQ
HYRPPFTFGQGTKLEIKR
S4-63
CDR-L1
Residues 24-34
KASQKVSSALA
VL
of SEQ ID
NO.: 775
S4-63
CDR-L2
Residues 50-56
WASALHS
VL
of SEQ ID
NO.: 775
S4-63
CDR-L3
Residues 89-97
QQHYRPPFT
VL
of SEQ ID
NO.: 775
S4-50
776
DIQMTQSPSSLSASVGDRVTITCKASQLVS
VL
SAVAWYQQKPGKAPKLLIYWASALHTGVPS
RFSGSGSGTDFTLTISSLQPEDFATYYCQQ
HYSSPYTFGQGTKLEIKR
S4-50
CDR-L1
Residues 24-34
KASQLVSSAVA
VL
of SEQ ID
NO.: 776
S4-50
CDR-L2
Residues 50-56
WASALHT
VL
of SEQ ID
NO.: 776
S4-50
CDR-L3
Residues 89-97
QQHYSSPYT
VL
of SEQ ID
NO.: 776
S4-19
777
DIQMTQSPSSLSASVGDRVTITCKASQLVS
VL
SAVAWYQQKPGKAPKLLIYWASTLHTGVPS
RFSGSGSGTDFTLTISSLQPEDFATYYCQQ
HYRTPFTFGQGTKLEIKR
S4-19
CDR-L1
Residues 24-34
KASQLVSSAVA
VL
of SEQ ID
NO.: 777
S4-19
CDR-L2
Residues 50-56
WASTLHT
VL
of SEQ ID
NO.: 777
S4-19
CDR-L3
Residues 89-97
QQHYRTPFT
VL
of SEQ ID
NO.: 777
TABLE 24
Heavy and light chain pairs of hMAK195 affinity matured clones
Clone name
HC
LC
Protein name
A8
hMAK195-A8
hMAK195 VL.1
hMAK195-AM11
B5
hMAK195-B5
hMAK195 VL.1
hMAK195-AM13
rHC3
hMAK195 rHC3
hMAK195 VL.1
hMAK195-AM14
rHC18
hMAK195 rHC18
hMAK195 VL.1
hMAK195-AM15
rHC19
hMAK195 rHC19
hMAK195 VL.1
hMAK195-AM16
rHC22
hMAK195 rHC22
hMAK195 VL.1
hMAK195-AM17
rHC34
hMAK195 rHC34
hMAK195 VL.1
hMAK195-AM18
rHC60
hMAK195 rHC60
hMAK195 VL.1
hMAK195-AM19
S4-6
hMAK195 S4-6
hMAK195 S4-6
hMAK195-AM20
S4-12
hMAK195 S4-12
hMAK195 S4-12
hMAK195-AM21
S4-17
hMAK195 S4-17
hMAK195 S4-17
hMAK195-AM22
S4-18
hMAK195 S4-18
hMAK195 S4-18
hMAK195-AM23
S4-19
hMAK195 S4-19
hMAK195 S4-19
hMAK195-AM24
S4-24
hMAK195 S4-24
hMAK195 S4-24
hMAK195-AM25
S4-34
hMAK195 S4-34
hMAK195 S4-34
hMAK195-AM26
TABLE 25
IgG Name
EC50 in hTNFa ELISA (nM)
hMAK195-AM11
0.2
hMAK195-AM13
0.2
hMAK195-AM14
0.051
hMAK195-AM15
0.052
hMAK195-AM16
0.056
hMAK195-AM17
0.056
hMAK195-AM18
0.052
hMAK195-AM19
0.057
hMAK195-AM20
0.043
hMAK195-AM21
0.042
hMAK195-AM22
0.052
hMAK195-AM23
0.055
hMAK195-AM24
0.053
hMAK195-AM25
0.052
hMAK195-AM26
0.061
TABLE 26
hu TNF neutralization
rhesus TNF neutralization
IgG Name
IC50 (nM)
IC50 (nM)
hMAK195-AM11
0.259
>25
hMAK195-AM13
1.218
4.64
hMAK195-AM14
0.0401
4.61
hMAK195-AM15
0.036
>150
hMAK195-AM16
0.0105
0.803
hMAK195-AM17
0.0031
>25
hMAK195-AM18
0.0145
0.4412
hMAK195-AM19
0.0126
1.206
hMAK195-AM20
0.0037
0.596
hMAK195-AM21
0.009
0.09
hMAK195-AM22
0.00345
0.2705
hMAK195-AM23
0.0468
2.627
hMAK195-AM24
0.015
0.557
hMAK195-AM25
0.0114
0.262
hMAK195-AM26
0.0061
0.2495
The mouse anti-human TNF antibody MAK-199 was humanized and affinity-matured to generate a panel of humanized MAK195 variants that have improved affinity and binding kinetics against both human and cyno TNF. Several libraries were made according to specifications below:
All four libraries were selected separately for the ability to bind human or cynomolgus monkey TNF in the presence of decreasing concentrations of biotinylated human or cynomolgus monkey TNF antigens. All mutated CDR sequences recovered from library selections were recombined into additional libraries and the recombined libraries were subjected to more stringent selection conditions before individual antibodies are identified.
Table 27 provides a list of amino acid sequences of VH of the hMAK-199 antibody which were subjected to the affinity maturation selection protocol Amino acid residues of individual CDRs of each VH sequence are indicated in bold.
TABLE 27
List of amino acid sequences of affinity matured hMAK199
VH variants
Clone
SEQ ID NO:
VH
J644M2S1-10VH
778
EVQLVQSGAEVKKPGASVKVSCKASGYTFNDYGITWVRQ
APGQGLEWMGWINTYTGKPTYAQGFTGRVTMTTDTSTST
AYMELSSLRSEDTAVYFCARKFLTTVVVTDYAMDYWGQG
TTVTVSS
J644M2S1-11VH
779
EVQLVQSGAEVKKPGASVKVSCKASGYTFTNYGMNWVRQ
APGQGLEWMGWINTYTGEPTYADDFKGRFTFTLDTSTST
AYMELSSLRSEDTAVYFCARKFLTTVVVTDYAMDYWGQG
TTVTVSS
J644M2S1-12VH
780
EVQLVQSGAEVKKPGASVKVSCKASGYTFTNYGISWVRQ
APGQGLEWMGWINTYTGEPHYAQGLTGRVTMTTDTSTST
AYMELSSLRSEDTAVYFCARKFLTTVVVTDYAMDYWGQG
TTVTVSS
J644M2S1-13VH
781
EVQLVQSGAEVKKPGASVKVSCKASGYTFDNYGIQWVRQ
APGQGLEWMGWINTYTGAPSYAQGFTGRVTMTTDTSTST
AYMELSSLRSEDTAVYFCARKFLTTVVVTDYAMDYWGQG
TTVTVSS
J644M2S1-14VH
782
EVQLVQSGAEVKKPGASVKVSCKASGYTFTDYGINWVRQ
APGQGLEWMGWINTYTGKPSYAQGFTGRVTMTTDTSTST
AYMELSSLRSEDTAVYFCARKFLTTVVVTDYAMDYWGQG
TTVTVSS
J644M2S1-15VH
783
EVQLVQSGAEVKKPGASVKVSCKASGYTFSNYGMNWVRQ
APGQGLEWMGWINTYTGESTYAQGFTGRVTMTTDTSTST
AYMELSSLRSEDTAVYFCARKFLTTVVVTDYAMDYWGQG
TTVTVSS
J644M2S1-16VH
784
EVQLVQSGAEVKKPGASVKVSCKASGYTFKNYGMTWVRQ
APGQGLEWMGWINTYTGKPTYAQGFTGRVTMTTDTSTST
AYMELSSLRSEDTAVYFCARKFLTTVVVTDYAMDYWGQG
TTVTVSS
J644M2S1-17VH
785
EVQLVQSGAEVKKPGASVKVSCKASGYAFTDYGINWVRQ
APGQGLEWMGWINTYTGKPTYAQGFTGRVTMTTDTSTST
AYMELSSLRSEDTAVYFCARKFLTTVVVTDYAMDYWGQG
TTVTVSS
J644M2S1-18VH
786
EVQLVQSGAEVKKPGASVKVSCKASGYTFNNYGINWVRQ
APGQGLEWMGWINTYTGEPAYAQGFTGRVTMTTDTSTST
AYMELSSLRSEDTAVYFCARKFLTTVVVTDYAMDYWGQG
TTVTVSS
J644M2S1-1VH
787
EVQLVQSGAEVKKPGASVKVSCKASGYTFRNYGINWVRQ
APGQGLEWMGWINTYTGQPTYAQGFTGRVTMTTDTSTST
AYMELSSLRSEDTAVYFCARKFLTTVVVTDYAMDYWGQG
TTVTVSS
J644M2S1-22VH
788
EVQLVQSGAEVKKPGASVKVSCKASGYTFTDYGINWVRQ
APGQGLEWMGWINTYTGEPTYAQGFTGRVTMTTDTSTST
AYMELSSLRSEDTAVYFCARKFLTTVVVTDYAMDYWGQG
TTVTVSS
J644M2S1-23VH
789
EVQLVQSGAEVKKPGASVKVSCKASGYTFKNYGIIWVRQ
APGQGLEWMGWINTYTGKPTYAQGFTGRVTMTTDTSTST
AYMELSSLRSEDTAVYFCARKFLTTVVVTDYAMDYWGQG
TTVTVSS
J644M2S1-24VH
790
EVQLVQSGAEVKKPGASVKVSCKASGYTFSNYGINWVRQ
APGQGLEWMGWINTYTGVPTYAQGFTGRVTMTTDTSTST
AYMELSSLRSEDTAVYFCARKFLTTVVVTDYAMDYWGQG
TTVTVSS
J644M2S1-25VH
791
EVQLVQSGAEVKKPGASVKVSCKASGYTFNNYGINWVRQ
APGQGLEWMGWINTYTGKPSYAQGFTGRVTMTTDTSTST
AYMELSSLRSEDTAVYFCARKFLTTVVVTDYAMDYWGQG
TTVTVSS
J644M2S1-27VH
792
EVQLVQSGAEVKKPGASVKVSCKASGYTFKNYGINWVRQ
APGQGLEWMGWINTYTGKPTYAQGLTGRVTMTTDTSTST
AYMELSSLRSEDTAVYFCARKFLTTVVVTDYAMDYWGQG
TTVTVSS
J644M2S1-28VH
793
EVQLVQSGAEVKKPGASVKVSCKASGYTFRNYGINWVRQ
APGQGLEWMGWINTYTGKPSYAQGFTGRVTMTTDTSTST
AYMELSSLRSEDTAVYFCARKFLTTVVVTDYAMDYWGQG
TTVTVSS
J644M2S1-2VH
794
EVQLVQSGAEVKKPGASVKVSCKASGYTFTNYGIXWVRQ
APGQGLEWMGWINTYXGKPTYAQGFTGRVTMTTDTSTST
AYMELSSLRSEDTAVYFCARKFLTTVVVTDYAMDYWGQG
TTVTVSS
J644M2S1-31VH
795
EVQLVQSGAEVKKPGASVKVSCKASGYTFNNYGINWVRQ
APGQGLEWMGWINTYTGEPHYAQGLTGRVTMTTDTSTST
AYMELSSLRSEDTAVYFCARKFLTTVVVTDYAMDYWGQG
TTVTVSS
J644M2S1-33VH
796
EVQLVQSGAEVKKPGASVKVSCKASGYTFTHYGINWVRQ
APGQGLEWMGWINTYTGKPTYAQGFTGRVTMTTDTSTST
AYMELSSLRSEDTAVYFCARKFLTTVVVTDYAMDYWGQG
TTVTVSS
J644M2S1-34VH
797
EVQLVQSGAEVKKPGASVKVSCKASGYTFTHYGINWVRQ
APGQGLEWMGWINTYTGQPTYAQGLTGRVTMTTDTSTST
AYMELSSLRSEDTAVYFCARKFLTTVVVTDYAMDYWGQG
TTVTVSS
J644M2S1-35VH
798
EVQLVQSGAEVKKPGASVKVSCKASGYTFTNYGITWVRQ
APGQGLEWMGWINTYTGKPTYAQGFTGRVTMTTDTSTST
AYMELSSLRSEDTAVYFCARKFLTTVVVTDYAMDYWGQG
TTVTVSS
J644M2S1-36VH
799
EVQLVQSGAEVKKPGASVKVSCKASGYTFGNYGINWVRQ
APGQGLEWMGWINTYTGKPSYAQGFTGRVTMTTDTSTST
AYMELSSLRSEDTAVYFCARKFLTTVVVTDYAMDYWGQG
TTVTVSS
J644M2S1-37VH
800
EVQLVQSGAEVKKPGASVKVSCKASGYTFSNYGINWVRQ
APGQGLEWMGWINTYTGRPTYAQGFTGRVTMTTDTSTST
AYMELSSLRSEDTAVYFCARKFLTTVVVTDYAMDYWGQG
TTVTVSS
J644M2S1-38VH
801
EVQLVQSGAEVKKPGASVKVSCKASGYTFKNYGINWVRQ
APGQGLEWMGWINTYTGEPHYAQGFTGRVTMTTDTSTST
AYIELSSLRSEDTAVYFCARKFLTTVVVTDYAMDYWGQG
TTVTVSS
J644M2S1-3VH
802
EVQLVQSGAEVKKPGASVKVSCKASGYTFTNYGINWVRQ
APGQGLEWMGWINTYTGEPSYAQGFTGRVTMTTDTSTST
AYMELSSLRSEDTAVYFCARKFLTTVVVTDYAMDYWGQG
TTVTVSS
J644M2S1-40VH
803
EVQLVQSGAEVKKPGASVKVSCKASGYTFSNYGMNWVRQ
APGQGLEWMGWINTYTGEPTYAQGFTGRVTMTTDTSTST
AYMELSSLRSEDTAVYFCARKFLTTVVVTDYAMDYWGQG
TTVTVSS
J644M2S1-41VH
804
EVQLVQSGAEVKKPGASVKVSCKASGYTFTNYGIGWVRQ
APGQGLEWMGWINTYTGKPSYAQGFTGRVTMTTDTSTST
AYMELSSLRSEDTAVYFCARKFLTTVVVTDYAMDYWGQG
TTVTVSS
J644M2S1-43VH
805
EVQLVQSGAEVKKPGASVKVSCKASGYTFNNYGINWVRQ
APGQGLEWMGWINTYTGVPSYAQGFTGRVTMTTDTSTST
AYMELSSLRSEDTAVYFCARKFLTTVVVTDYAMDYWGQG
TTVTVSS
J644M2S1-44VH
806
EVQLVQSGAEVKKPGASVKVSCKASGYTFSNYGIAWVRQ
APGQGLEWMGWINTYTGVPTYAQGFTGRVTMTTDTSTST
AYMELSSLRSEDTAVYFCARKFLTTVVVTDYAMDYWGQG
TTVTVSS
J644M2S1-45VH
807
EVQLVQSGAEVKKPGASVKVSCKASGYTFNNYGINWVRQ
APGQGLEWMGWINTYTGVPHYAQGFTGRVTMTTDTSTST
AYMELSSLRSEDTAVYFCARKFLTTVVVTDYAMDYWGQG
TTVTVSS
J644M2S1-46VH
808
EVQLVQSGAEVKKPGASVKVSCKASGYTFTNYGIXWVRQ
APGQGLEWMGWINTYTGEPXYAQGFTGRVTMTTDTSTST
AYMELSSLRSEDTAVYFCARKFLTTVVVTDYAMDYWGQG
TTVTVSS
J644M2S1-47VH
809
EVQLVQSGAEVKKPGASVKVSCKASGYTFNNYGINWVRQ
APGQGLEWMGWINTYTGVPTYAQGLTGRVTMTTDTSTST
AYMELSSLRSEDTAVYFCARKFLTTVVVTDYAMDYWGQG
TTVTVSS
J644M2S1-48VH
810
EVQLVQSGAEVKKPGASVKVSCKASGYTFTNYGINWVRQ
APGQGLEWMGWINTYTGQPTYAQGFTGRVTMTTDTSTST
AYMELSSLRSEDTAVYFCARKFLTTVVVTDYAMDYWGQG
TTVTVSS
J644M2S1-4VH
811
EVQLVQSGAEVKKPGASVKVSCKASGYTFTDYGITWVRQ
APGQGLEWMGWINTYTGKPTYAQGFTGRVTMTTDTSTST
AYMELSSLRSEDTAVYFCARKFLTTVVVTDYAMDYWGQG
TTVTVSS
J644M2S1-50VH
812
EVQLVQSGAEVKKPGASVKVSCKASGYTFTNYGINWVRQ
APGQGLEWMGWINTYTGVPQYAQGFTGRVTMTTDTSTST
AYMELSSLRSEDTAVYFCARKFLTTVVVTDYAMDYWGQG
TTVTVSS
J644M2S1-51VH
813
EVQLVQSGAEVKKPGASVKVSCKASGYTFQNYGINWVRQ
APGQGLEWMGWINTYTGVPTYAQGLTGRVTMTTDTSTST
AYMELSSLRSEDTAVYFCARKFLTTVVVTDYAMDYWGQG
TTVTVSS
J644M2S1-53VH
814
EVQLVQSGAEVKKPGASVKVSCKASGYTFTQYGINWVRQ
APGQGLEWMGWINTYTGDPHYAQGFTGRVTMTTDTSTST
AYMELSSLRSEDTAVYFCARKFLTTVVVTDYAMDYWGQG
TTVTVSS
J644M2S1-54VH
815
EVQLVQSGAEVKKPGASVKVSCKASGYTFSNYGINWVRQ
APGQGLEWMGWINTYTGLPTYAQGFTGRVTMTTDTSTST
AYMELSSLRSEDTAVYFCARKFLTTVVVTDYAMDYWGQG
TTVTVSS
J644M2S1-55VH
816
EVQLVQSGAEVKKPGASVKVSCKASGYTFTNYGMNWVRQ
APGQGLEWMGWINTYNGKPMYAQGFTGRVTMTTDTSTST
AYMELSSLRSEDTAVYFCARKFLTTVVVTDYAMDYWGQG
TTVTVSS
J644M2S1-56VH
817
EVQLVQSGAEVKKPGASVKVSCKASGYTFRNYGITWVRQ
APGQGLEWMGWINTYTGEPAYAQGLTGRVTMTTDTSTST
AYMELSSLRSEDTAVYFCARKFLTTVVVTDYAMDYWGQG
TTVTVSS
J644M2S1-59VH
818
EVQLVQSGAEVKKPGASVKVSCKASGYTFNHYGINWVRQ
APGQGLEWMGWINTYTGRPTYAQGFTGRVTMTTDTSTST
AYMELSSLRSEDTAVYFCARKFLTTVVVTDYAMDYWGQG
TTVTVSS
J644M2S1-5VH
819
EVQLVQSGAEVKKPGASVKVSCKASGYTFSNYGINWVRQ
APGQGLEWMGWINTYTGKPTYAQGLTGRVTMTTDTSTST
AYMELSSLRSEDTAVYFCARKFLTTVVVTDYAMDYWGQG
TTVTVSS
J644M2S1-60VH
820
EVQLVQSGAEVKKPGASVKVSCKASGYTFNNYGINWVRQ
APGQGLEWMGWINTYTGKPTYAQGFTGRVTMTTDTSTST
AYMELSSLRSEDTAVYFCARKFLTTVVVTDYAMDYWGQG
TTVTVSS
J644M2S1-64VH
821
EVQLVQSGAEVKKPGASVKVSCKASGYTFDNYGINWVRQ
APGQGLEWMGWINTYTGVPTYAQGFTGRVTMTTDTSTST
AYMELSSLRSEDTAVYFCARKFLTTVVVTDYAMDYWGQG
TTVTVSS
J644M2S1-65VH
822
EVQLVQSGAEVKKPGASVKVSCKASGYTFNDYGIIWVRQ
APGQGLEWMGWINTYTGKPSYAQGFTGRVTMTTDTSTST
AYMELSSLRSEDTAVYFCARKFLTTVVVTDYAMDYWGQG
TTVTVSS
J644M2S1-66VH
823
EVQLVQSGAEVKKPGASVKVSCKASGYTFSNYGINWVRQ
APGQGLEWMGWINTYTGKPTYAQGFTGRVTMTTDTSTST
AYMELSSLRSEDTAVYFCARKFLTTVVVTDYAMDYWGQG
TTVTVSS
J644M2S1-67VH
824
EVQLVQSGAEVKKPGASVKVSCKASGYTFANYGMNWVRQ
APGQGLEWMGWINTYTGKPTYAQGFTGRVTMTTDTSTST
AYMELSSLRSEDTAVYFCARKFLTTVVVTDYAMDYWGQG
TTVTVSS
J644M2S1-68VH
825
EVQLVQSGAEVKKPGASVKVSCKASGYTFNNYGINWVRQ
APGQGLEWMGWINTYTGEPSYAQGLTGRVTMTTDTSTST
AYMELSSLRSEDTAVYFCARKFLTTVVVTDYAMDYWGQG
TTVTVSS
J644M2S1-6VH
826
EVQLVQSGAEVKKPGASVKVSCKASGYTFTNYGINWVRQ
APGQGLEWMGWINTYTGVPTYAQGLTGRVTMTTDTSTST
AYMELSSLRSEDTAVYFCARKFLTTVVVTDYAMDYWGQG
TTVTVSS
J644M2S1-71VH
827
EVQLVQSGAEVKKPGASVKVSCKASGYTFDHYGMNWVRQ
APGQGLEWMGWINTYTGKPTYAQGLTGRVTMTTDTSTST
AYMELSSLRSEDTAVYFCARKFLTTVVVTDYAMDYWGQG
TTVTVSS
J644M2S1-72VH
828
EVQLVQSGAEVKKPGASVKVSCKASGYTFTNYGIGWVRQ
APGQGLEWMGWINTYTGKPSYAQGLTGRVTMTTDTSTST
AYMELSSLRSEDTAVYFCARKFLTTVVVTDYAMDYWGQG
TTVTVSS
J644M2S1-73VH
829
EVQLVQSGAEVKKPGASVKVSCKASGYTFTNYGMNWVRQ
APGQGLEWMGWINTYTGEPTYADDFKGRFTFTLDTSTST
AYMELSSLRSEDTAVYFCARKFLTTVVVTDYAMDYWGQG
TTVTVSS
J644M2S1-74VH
830
EVQLVQSGAEVKKPGASVKVSCKASGYTFNNYGMNWVRQ
APGQGLEWMGWINTYTGKPTYAQGLTGRVTMTTDTSTST
AYMELSSLRSEDTAVYFCARKFLTTVVVTDYAMDYWGQG
TTVTVSS
J644M2S1-75VH
831
EVQLVQSGAEVKKPGASVKVSCKASGYTFDNYGMNWVRQ
APGQGLEWMGWINTYTGVPTYAQGFTGRVTMTTDTSTST
AYMELSSLRSEDTAVYFCARKFLTTVVVTDYAMDYWGQG
TTVTVSS
J644M2S1-76VH
832
EVQLVQSGAEVKKPGASVKVSCKASGYTFNSYGINWVRQ
APGQGLEWMGWINTYTGKPTYAQGFTGRVTMTTDTSTST
AYMELSSLRSEDTAVYFCARKFLTTVVVTDYAMDYWGQG
TTVTVSS
J644M2S1-77VH
833
EVQLVQSGAEVKKPGASVKVSCKASGYTFSNYGITWVRQ
APGQGLEWMGWINTYTGKPTYAQGFTGRVTMTTDTSTST
AYMELSSLRSEDTAVYFCARKFLTTVVVTDYAMDYWGQG
TTVTVSS
J644M2S1-79VH
834
EVQLVQSGAEVKKPGASVKVSCKASGYTFTNYGINWVRQ
APGQGLEWMGWINTYNGQPTYAQGFTGRVTMTTDTSTST
AYMELSSLRSEDTAVYFCARKFLTTVVVTDYAMDYWGQG
TTVTVSS
J644M2S1-7VH
835
EVQLVQSGAEVKKPGASVKVSCKASGYTFTNYGIIWVRQ
APGQGLEWMGWINTYTGEPTYAQGFTGRVTMTTDTSTST
AYMELSSLRSEDTAVYFCARKFLTTVVVTDYAMDYWGQG
TTVTVSS
J644M2S1-81VH
836
EVQLVQSGAEVKKPGASVKVSCKASGYTFANYGINWVRQ
APGQGLEWMGWINTYTGKPTYAQGFTGRVTMTTDTSTST
AYMELSSLRSEDTAVYFCARKFLTTVVVTDYAMDYWGQG
TTVTVSS
J644M2S1-82VH
837
EVQLVQSGAEVKKPGASVKVSCKASGYTFSDYGIQWVRQ
APGQGLEWMGWINTYTGRPTYAQGFTGRVTMTTDTSTST
AYMELSSLRSEDTAVYFCARKFLTTVVVTDYAMDYWGQG
TTVTVSS
J644M2S1-83VH
838
EVQLVQSGAEVKKPGASVKVSCKASGYTFSNYGISWVRQ
APGQGLEWMGWINTYTGKPTYAQGFTGRVTMTTDTSTST
AYMELSSLRSEDTAVYFCARKFLTTVVVTDYAMDYWGQG
TTVTVSS
J644M2S1-84VH
839
EVQLVQSGAEVKKPGASVKVSCKASGYTFTNYGIQWVRQ
APGQGLEWMGWINTYTGVPTYAQGFTGRVTMTTDTSTST
AYMELSSLRSEDTAVYFCARKFLTTVVVTDYAMDYWGQG
TTVTVSS
J644M2S1-85VH
840
EVQLVQSGAEVKKPGASVKVSCKASGYTFTNYGINWVRQ
APGQGLEWMGWINTYTGVPTYAQGFTGRVTMTTDTSTST
AYMELSSLRSEDTAVYFCARKFLTTVVVTDYAMDYWGQG
TTVTVSS
J644M2S1-87VH
841
EVQLVQSGAEVKKPGASVKVSCKASGYTFTNYGINWVRQ
APGQGLEWMGWINTYSGKPTYAQGFTGRVTMTTDTSTST
AYMELSSLRSEDTAVYFCARKFLTTVVVTDYAMDYWGQG
TTVTVSS
J644M2S1-88VH
842
EVQLVQSGAEVKKPGASVKVSCKASGYTFSNYGINWVRQ
APGQGLEWMGWINTYTGQPTYAQGFTGRVTMTTDTSTST
AYMELSSLRSEDTAVYFCARKFLTTVVVTDYAMDYWGQG
TTVTVSS
J644M2S1-8VH
843
EVQLVQSGAEVKKPGASVKVSCKASGYTFPNYGINWVRQ
APGQGLEWMGWINTYTGKPTYAQGFTGRVTMTTDTSTST
AYMELSSLRSEDTAVYFCARKFLTTVVVTDYAMDYWGQG
TTVTVSS
J644M2S1-90VH
844
EVQLVQSGAEVKKPGASVKVSCKASGYTFTNYGINWVRQ
APGQGLEWMGWINTYTGKTNYAQGFTGRVTMTTDTSTST
AYMELSSLRSEDTAVYFCARKFLTTVVVTDYAMDYWGQG
TTVTVSS
J644M2S1-91VH
845
EVQLVQSGAEVKKPGASVKVSCKASGYTFNNYGINWVRQ
APGQGLEWMGWINTYTGEPNYAQGFTGRVTMTTDTSTST
AYMELSSLRSEDTAVYFCARKFLTTVVVTDYAMDYWGQG
TTVTVSS
J644M2S1-92VH
846
EVQLVQSGAEVKKPGASVKVSCKASGYTFSNYGITWVRQ
APGQGLEWMGWINTYTGEPHYAQGLTGRVTMTTDTSTST
AYMELSSLRSEDTAVYFCARKFLTTVVVTDYAMDYWGQG
TTVTVSS
J644M2S1-93VH
847
EVQLVQSGAEVKKPGASVKVSCKASGYTFKNYGINWVRQ
APGQGLEWMGWINTYTGQPTYAQGFTGRVTMTTDTSTST
AYMELSSLRSEDTAVYFCARKFLTTVVVTDYAMDYWGQG
TTVTVSS
J644M2S1-94VH
848
EVQLVQSGAEVKKPGASVKVSCKASGYTFNNYGINWVRQ
APGQGLEWMGWINTYTGIPTYAQGFTGRVTMTTDTSTST
AYMELSSLRSEDTAVYFCARKFLTTVVVTDYAMDYWGQG
TTVTVSS
J644M2S1-95VH
849
EVQLVQSGAEVKKPGASVKVSCKASGYTFTDYGINWVRQ
APGQGLEWMGWINTYTGKPTYAQGFTGRVTMTTDTSTST
AYMELSSLRSEDTAVYFCARKFLTTVVVTDYAMDYWGQG
TTVTVSS
J644M2S1-96VH
850
EVQLVQSGAEVKKPGASVKVSCKASGYTFSNYGINWVRQ
APGQGLEWMGWINTYSGVPTYAQGFTGRVTMTTDTSTST
AYMELSSLRSEDTAVYFCARKFLTTVVVTDYAMDYWGQG
TTVTVSS
J644M2S1-9VH
851
EVQLVQSGAEVKKPGASVKVSCKASGYTFTDYGINWVRQ
APGQGLEWMGWINTYTGKPTYAQGFTGRVTMTTDTSTST
AYMELSSLRSEDTAVYFCARKFLTTVVVTDYAMDYWGQG
TTVTVSS
J647M2-11VH
852
EVQLVQSGAEVKKPGASVKVSCKASGYTFTNYGMNWVRQ
APGQGLEWMGWINTYTGEPTYADDFKGRFTFTLDTSTST
AYMELSSLRSEDTAVYYCARKFWRTVVGTDNAMDYWGQG
TTVTVSS
J647M2-12VH
853
EVQLVQSGAEVKKPGASVKVSCKASGYTFTNYGMNWVRQ
APGQGLEWMGWINTYTGEPTYADDFKGRFTFTLDTSTST
AYMELSSLRSEDTAVYFCARKYSTTVVVTDYAMDYWGQG
TTVTVSS
J647M2-13VH
854
EVQLVQSGAEVKKPGASVKVSCKASGYTFTNYGMNWVRQ
APGQGLEWMGWINTYTGEPTYADDFKGRFTFTLDTSTST
AYMELSSLRSEDTAVYFCARKFLTTVVVTDSAMDYWGQG
TTVTVSS
J647M2-15VH
855
EVQLVQSGAEVKKPGASVKVSCKASGYTFTNYGMNWVRQ
APGQGLEWMGWINTYTGEPTYADDFKGRFTFTLDTSTST
AYMELSSLRSEDTAVYFCARKFMTTMAVTDFAMDYWGQG
TTVTVSS
J647M2-16VH
856
EVQLVQSGAEVKKPGASVKVSCKASGYTFTNYGMNWVRQ
APGQGLEWMGWINTYTGEPTYADDFKGRFTFTLDTSTST
AYMELSSLRSEDTAVYYCARKLLTTVVATDNAMDYWGQG
TTVTVSS
J647M2-17VH
857
EVQLVQSGAEVKKPGASVKVSCKASGYTFTNYGMNWVRQ
APGQGLEWMGWINTYTGEPTYADDFKGRFTFTLDTSTST
AYMELSSLRSEDTAVYYCARKFLTTVIVTDNAMDYWGQG
TTVTVSS
J647M2-19VH
858
EVQLVQSGAEVKKPGASVKVSCKASGYTFTNYGMNWVRQ
APGQGLEWMGWINTYTGEPTYADDFKGRFTFTLDTSTST
AYMELSSLRSEDTAVYFCARKFFTPVVVTDNAMDYWGQG
TTVTVSS
J647M2-1VH
859
EVQLVQSGAEVKKPGASVKVSCKASGYTFTNYGMNWVRQ
APGQGLEWMGWINTYTGEPTYADDFKGRFTFTLDTSTST
AYMELSSLRSEDTAVYYCARKLMTTVVVTDHAMDYWGQG
TTVTVSS
J647M2-20VH
860
EVQLVQSGAEVKKPGASVKVSCKASGYTFTNYGMNWVRQ
APGQGLEWMGWINTYTGEPTYADDFKGRFTFTLDTSTST
AYMELSSLRSEDTAVYYCARKYLTTVVVTDSAMDYWGQG
TTVTVSS
J647M2-21VH
861
EVQLVQSGAEVKKPGASVKVSCKASGYTFTNYGMNWVRQ
APGQGLEWMGWINTYTGEPTYADDFKGRFTFTLDTSTST
AYMELSSLRSEDTAVYYCARKFRSSVAVTDNAMDYWGQG
TTVTVSS
J647M2-22VH
862
EVQLVQSGAEVKKPGASVKVSCKASGYTFTNYGMNWVRQ
APGQGLEWMGWINTYTGEPTYADDFKGRFTFTLDTSTST
AYMELSSLRSEDTAVYYCARKLFTTVVVTDSAMDYWGQG
TTVTVSS
J647M2-23VH
863
EVQLVQSGAEVKKPGASVKVSCKASGYTFTNYGMNWVRQ
APGQGLEWMGWINTYTGEPTYADDFKGRFTFTLDTSTST
AYMELSSLRSEDTAVYFCARKYLMPVVVTDYAMDYWGQG
TTVTVSS
J647M2-24VH
864
EVQLVQSGAEVKKPGASVKVSCKASGYTFTNYGMNWVRQ
APGQGLEWMGWINTYTGEPTYADDFKGRFTFTLDTSTST
AYMELSSLRSEDTAVYFCARKLLDAVMVTDYAMDYWGQG
TTVTVSS
J647M2-26VH
865
EVQLVQSGAEVKKPGASVKVSCKASGYTFTNYGMNWVRQ
APGQGLEWMGWINTYTGEPTYADDFKGRFTFTLDTSTST
AYMELSSLRSEDTAVYYCARKFLTTVVVNDYAMDYWGQG
TTVTVSS
J647M2-44VH
866
EVQLVQSGAEVKKPGASVKVSCKASGYTFTNYGMNWVRQ
APGQGLEWMGWINTYTGEPTYADDFKGRFTFTLDTSTST
AYMELSSLRSEDTAVYYCARKLLTTVAVTDYAMDYWGQG
TTVTVSS
J647M2-45VH
867
EVQLVQSGAEVKKPGASVKVSCKASGYTFTNYGMNWVRQ
APGQGLEWMGWINTYTGEPTYADDFKGRFTFTLDTSTST
AYMELSSLRSEDTAVYFCARKFLKTVVATDDAMDYWGQG
TTVTVSS
J647M2-47VH
868
EVQLVQSGAEVKKPGASVKVSCKASGYTFTNYGMNWVRQ
APGQGLEWMGWINTYTGEPTYADDFKGRFTFTLDTSTST
AYMELSSLRSEDTAVYYCARKFLNTAVVTDYAMDYWGQG
TTVTVSS
J647M2-48VH
869
EVQLVQSGAEVKKPGASVKVSCKASGYTFTNYGMNWVRQ
APGQGLEWMGWINTYTGEPTYADDFKGRFTFTLDTSTST
AYMELSSLRSEDTAVYYCARRFLTTVDVTDNAMDYWGQG
TTVTVSS
J647M2-4VH
870
EVQLVQSGAEVKKPGASVKVSCKASGYTFTNYGMNWVRQ
APGQGLEWMGWINTYTGEPTYADDFKGRFTFTLDTSTST
AYMELSSLRSEDTAVYFCARKYLTPVVATDFAMDYWGQG
TTVTVSS
J647M2-51VH
871
EVQLVQSGAEVKKPGASVKVSCKASGYTFTNYGMNWVRQ
APGQGLEWMGWINTYTGEPTYADDFKGRFTFTLDTSTST
AYMELSSLRSEDTAVYFCARKCMTTIVETDNAMDYWGQG
TTVTVSS
J647M2-52VH
872
EVQLVQSGAEVKKPGASVKVSCKASGYTFTNYGMNWVRQ
APGQGLEWMGWINTYTGEPTYADDFKGRFTFTLDTSTST
AYMELSSLRSEDTAVYYCARKFMNTVDVTDNAMDYWGQG
TTVTVSS
J647M2-53VH
873
EVQLVQSGAEVKKPGASVKVSCKASGYTFTNYGMNWVRQ
APGQGLEWMGWINTYTGEPTYADDFKGRFTFTLDTSTST
AYMELSSLRSEDTAVYYCARKLFTTVVVTDDAMDYWGQG
TTVTVSS
J647M2-54VH
874
EVQLVQSGAEVKKPGASVKVSCKASGYTFTNYGMNWVRQ
APGQGLEWMGWINTYTGEPTYADDFKGRFTFTLDTSTST
AYMELSSLRSEDTAVYYCARKLMTTVVVTDYAMDYWGQG
TTVTVSS
J647M2-55VH
875
EVQLVQSGAEVKKPGASVKVSCKASGYTFTNYGMNWVRQ
APGQGLEWMGWINTYTGEPTYADDFKGRFTFTLDTSTST
AYMELSSLRSEDTAVYYCARKFLPTVVVTDYAMDYWGQG
TTVTVSS
J647M2-56VH
876
EVQLVQSGAEVKKPGASVKVSCKASGYTFTNYGMNWVRQ
APGQGLEWMGWINTYTGEPTYADDFKGRFTFTLDTSTST
AYMELSSLRSEDTAVYFCARKLLTTVVVTDNAMDYWGQG
TTVTVSS
J647M2-58VH
877
EVQLVQSGAEVKKPGASVKVSCKASGYTFTNYGMNWVRQ
APGQGLEWMGWINTYTGEPTYADDFKGRFTFTLDTSTST
AYMELSSLRSEDTAVYYCARKILTTVVVTDNAMDYWGQG
TTVTVSS
J647M2-70VH
878
EVQLVQSGAEVKKPGASVKVSCKASGYTFTNYGMNWVRQ
APGQGLEWMGWINTYTGEPTYADDFKGRFTFTLDTSTST
AYMELSSLRSEDTAVYYCARKVMATEVVTDYAMDYWGQG
TTVTVSS
J647M2-71VH
879
EVQLVQSGAEVKKPGASVKVSCKASGYTFTNYGMNWVRQ
APGQGLEWMGWINTYTGEPTYADDFKGRFTFTLDTSTST
AYMELSSLRSEDTAVYYCARKLVTTVVVTDYAMDYWGQG
TTVTVSS
J647M2-72VH
880
EVQLVQSGAEVKKPGASVKVSCKASGYTFTNYGMNWVRQ
APGQGLEWMGWINTYTGEPTYADDFKGRFTFTLDTSTST
AYMELSSLRSEDTAVYFCARKFRKPVSVTDYAMDYWGQG
TTVTVSS
J647M2-73VH
881
EVQLVQSGAEVKKPGASVKVSCKASGYTFTNYGMNWVRQ
APGQGLEWMGWINTYTGEPTYADDFKGRFTFTLDTSTST
AYMELSSLRSEDTAVYYCARKLWTTVVVTDNAMDYWGQG
TTVTVSS
J647M2-74VH
882
EVQLVQSGAEVKKPGASVKVSCKASGYTFTNYGMNWVRQ
APGQGLEWMGWINTYTGEPTYADDFKGRFTFTLDTSTST
AYMELSSLRSEDTAVYYCARKLLTPVVVTDYAMDYWGQG
TTVTVSS
J647M2-75VH
883
EVQLVQSGAEVKKPGASVKVSCKASGYTFTNYGMNWVRQ
APGQGLEWMGWINTYTGEPTYADDFKGRFTFTLDTSTST
AYMELSSLRSEDTAVYYCARKFRTTVVETDYCMDYWGQG
TTVTVSS
J647M2-76VH
884
EVQLVQSGAEVKKPGASVKVSCKASGYTFTNYGMNWVRQ
APGQGLEWMGWINTYTGEPTYADDFKGRFTFTLDTSTST
AYMELSSLRSEDTAVYFCARKYFTTVAVTDYAMDYWGQG
TTVTVSS
J647M2-78VH
885
EVQLVQSGAEVKKPGASVKVSCKASGYTFTNYGMNWVRQ
APGQGLEWMGWINTYTGEPTYADDFKGRFTFTLDTSTST
AYMELSSLRSEDTAVYYCARRFLTTVEVTDLAMDYWGQG
TTVTVSS
J647M2-79VH
886
EVQLVQSGAEVKKPGASVKVSCKASGYTFTNYGMNWVRQ
APGQGLEWMGWINTYTGEPTYADDFKGRFTFTLDTSTST
AYMELSSLRSEDTAVYYCARKFLRTEVMTDYAMDYWGQG
TTVTVSS
J647M2-7VH
887
EVQLVQSGAEVKKPGASVKVSCKASGYTFTNYGMNWVRQ
APGQGLEWMGWINTYTGEPTYADDFKGRFTFTLDTSTST
AYMELSSLRSEDTAVYFCARKFLSTVAVTDSAMDYWGQG
TTVTVSS
J647M2-80VH
888
EVQLVQSGAEVKKPGASVKVSCKASGYTFTNYGMNWVRQ
APGQGLEWMGWINTYTGEPTYADDFKGRFTFTLDTSTST
AYMELSSLRSEDTAVYYCARKVLNTVVVTDYAMDYWGQG
TTVTVSS
J647M2-83VH
889
EVQLVQSGAEVKKPGASVKVSCKASGYTFTNYGMNWVRQ
APGQGLEWMGWINTYTGEPTYADDFKGRFTFTLDTSTST
AYMELSSLRSEDTAVYFCARKFMNTAMVTDYAMDYWGQG
TTVTVSS
J647M2-84VH
890
EVQLVQSGAEVKKPGASVKVSCKASGYTFTNYGMNWVRQ
APGQGLEWMGWINTYTGEPTYADDFKGRFTFTLDTSTST
AYMELSSLRSEDTAVYYCARKFSTTVVVTDYAMDYWGQG
TTVTVSS
J647M2-85VH
891
EVQLVQSGAEVKKPGASVKVSCKASGYTFTNYGMNWVRQ
APGQGLEWMGWINTYTGEPTYADDFKGRFTFTLDTSTST
AYMELSSLRSEDTAVYFCARKYFTTVVVTDYAMDYWGQG
TTVTVSS
J647M2-86VH
892
EVQLVQSGAEVKKPGASVKVSCKASGYTFTNYGMNWVRQ
APGQGLEWMGWINTYTGEPTYADDFKGRFTFTLDTSTST
AYMELSSLRSEDTAVYFCARKFLNTVVVTDYAMDYWGQG
TTVTVSS
J647M2S1-12VH
893
EVQLVQSGAEVKKPGASVKVSCKASGYTFTNYGMNWVRQ
APGQGLEWMGWINTYTGEPTYADDFKGRFTFTLDTSTST
AYMELSSLRSEDTAVYYCARKFMPTVVETDYAMDYWGQG
TTVTVSS
J647M2S1-13VH
894
EVQLVQSGAEVKKPGASVKVSCKASGYTFTNYGINWVRQ
APGQGLEWMGWINTYTGNPTYAQGFTGRVTMTTDTSTST
AYMELSSLRSEDTAVYFCARKFLTTVVVTDYAMDYWGQG
TTVTVSS
J647M2S1-14VH
895
EVQLVQSGAEVKKPGASVKVSCKASGYTFADYGMNWVRQ
APGQGLEWMGWINTYTGEPTYAQGFTGRVTMTTDTSTST
AYMELSSLRSEDTAVYFCARKFLTTVVVTDYAMDYWGQG
TTVTVSS
J647M2S1-15VH
896
EVQLVQSGAEVKKPGASVKVSCKASGYTFTNYGMNWVRQ
APGQGLEWMGWINTYTGEPTYADDFKGRFTFTLDTSTST
AYMELSSLRSEDTAVYYCARKFLTTVVVTDCAMDYWGQG
TTVTVSS
J647M2S1-17VH
897
EVQLVQSGAEVKKPGASVKVSCKASGYTFTNYGMNWVRQ
APGQGLEWMGWINTYTGEPTYAQGFTGRVTMTTDTSTST
AYMELSSLRSEDTAVYFCARKFLTTVVVTDYAMDYWGQG
TTVTVSS
J647M2S1-18VH
898
EVQLVQSGAEVKKPGASVKVSCKASGYTFTNYGMNWVRQ
APGQGLEWMGWINTYTGEPTYADDFKGRFTFTLDTSTST
AYMELSSLRSEDTAVYYCARKFLTTVVVTDNAMDYWGQG
TTVTVSS
J647M2S1-19VH
899
EVQLVQSGAEVKKPGASVKVSCKASGYTFTNYGMNWVRQ
APGQGLEWMGWINTYTGEPTYADDFKGRFTFTLDTSTST
AYMELSSLRSEDTAVYYCARKLLNTVVGTDYAMDYWGQG
TTVTVSS
J647M2S1-21VH
900
EVQLVQSGAEVKKPGASVKVSCKASGYTFTNYGMNWVRQ
APGQGLEWMGWINTYTGEPTYADDFKGRFTFTLDTSTST
AYMELSSLRSEDTAVYFCARKLLTTEAVTDYAMDYWGQG
TTVTVSS
J647M2S1-22VH
901
EVQLVQSGAEVKKPGASVKVSCKASGYTFTNYGMNWVRQ
APGQGLEWMGWINTYTGEPTYADDFKGRFTFTLDTSTST
AYMELSSLRSEDTAVYFCARKYSTPVVVTDYAMDYWGQG
TTVTVSS
J647M2S1-23VH
902
EVQLVQSGAEVKKPGASVKVSCKASGYTFTNYGINWVRQ
APGQGLEWMGWINTYTGEPTYAQGLTGRVTMTTDTSTST
AYMELSSLRSEDTAVYFCARKFLTTVVVTDYAMDYWGQG
TTVTVSS
J647M2S1-26VH
903
EVQLVQSGAEVKKPGASVKVSCKASGYTFTNYGMNWVRQ
APGQGLEWMGWINTYTGEPTYADDFKGRFTFTLDTSTST
AYMELSSLRSEDTAVYFCARKCLNTVAVTEHRMDYWGQG
TTVTVSS
J647M2S1-28VH
904
EVQLVQSGAEVKKPGASVKVSCKASGYTFTNYGMNWVRQ
APGQGLEWMGWINTYTGEPTYADDFKGRFTFTLDTSTST
AYMELSSLRSEDTAVYYCARKFLTTVVHTDYAMDYWGQG
TTVTVSS
J647M2S1-30VH
905
EVQLVQSGAEVKKPGASVKVSCKASGYTFTNYGMNWVRQ
APGQGLEWMGWINTYTGQPTYAQGFTGRVTMTTDTSTST
AYMELSSLRSEDTAVYFCARKFLTTVVVTDYAMDYWGQG
TTVTVSS
J647M2S1-31VH
906
EVQLVQSGAEVKKPGASVKVSCKASGYTFTNYGINWVRQ
APGQGLEWMGWINTYTGKPTYAQGFTGRVTMTTDTSTST
AYMELSSLRSEDTAVYFCARKFLTTVVVTDYAMDYWGQG
TTVTVSS
J647M2S1-32VH
907
EVQLVQSGAEVKKPGASVKVSCKASGYTFANYGINWVRQ
APGQGLEWMGWINTYTGEPTYAQGFTGRVTMTTDTSTST
AYMELSSLRSEDTAVYFCARKFLTTVVVTDYAMDYWGQG
TTVTVSS
J647M2S1-33VH
908
EVQLVQSGAEVKKPGASVKVSCKASGYTFTNYGMNWVRQ
APGQGLEWMGWINTYTGEPTYADDFKGRFTFTLDTSTST
AYMELSSLRSEDTAVYYCARKFRTTVVLTDSAMDYWGQG
TTVTVSS
J647M2S1-35VH
909
EVQLVQSGAEVKKPGASVKVSCKASGYTFSNYGINWVRQ
APGQGLEWMGWINTYTGEPTYAQGLTGRVTMTTDTSTST
AYMELSSLRSEDTAVYFCARKFLTTVVVTDYAMDYWGQG
TTVTVSS
J647M2S1-36VH
910
EVQLVQSGAEVKKPGASVKVSCKASGYTFTNYGMNWVRQ
APGQGLEWMGWINTYTGEPTYADDFKGRFTFTLDTSTST
AYMELSSLRSEDTAVYYCARKFQTPVVDTDYAMDYWGQG
TTVTVSS
J647M2S1-39VH
911
EVQLVQSGAEVKKPGASVKVSCKASGYTFTNYGMNWVRQ
APGQGLEWMGWINTYTGEPTYADDFKGRFTFTLDTSTST
AYMELSSLRSEDTAVYYCARKFMKTRVVTDNAMDYWGQG
TTVTVSS
J647M2S1-40VH
912
EVQLVQSGAEVKKPGASVKVSCKASGYTFSNYGIVWVRQ
APGQGLEWMGWINTYTGEPTYAQGFTGRVTMTTDTSTST
AYMELSSLRSEDTAVYFCARKFLTTVVVTDYAMDYWGQG
TTVTVSS
J647M2S1-41VH
913
EVQLVQSGAEVKKPGASVKVSCKASGYTFPNYGISWVRQ
APGQGLEWMGWINTYTGEPSYAQGFTGRVTMTTDTSTST
AYMELSSLRSEDTAVYFCARKFLTTVVVTDYAMDYWGQG
TTVTVSS
J647M2S1-43VH
914
EVQLVQSGAEVKKPGASVKVSCKASGYTFTNYGINWVRQ
APGQGLEWMGWINTYTGEPSYAQGLTGRVTMTTDTSTST
AYMELSSLRSEDTAVYFCARKFLTTVVVTDYAMDYWGQG
TTVTVSS
J647M2S1-45VH
915
EVQLVQSGAEVKKPGASVKVSCKASGYTFTKYGINWVRQ
APGQGLEWMGWINTYTGEPTYAQGFTGRVTMTTDTSTST
AYMELSSLRSEDTAVYFCARKFLTTVVVTDYAMDYWGQG
TTVTVSS
J647M2S1-47VH
916
EVQLVQSGAEVKKPGASVKVSCKASGYTFTNYGMNWVRQ
APGQGLEWMGWINTYTGEPTYADDFKGRFTFTLDTSTST
AYMELSSLRSEDTAVYYCARKYLTTVVATDYAMDYWGQG
TTVTVSS
J647M2S1-48VH
917
EVQLVQSGAEVKKPGASVKVSCKASGYTFTNYGMNWVRQ
APGQGLEWMGWINTYTGEPTYADDFKGRFTFTLDTSTST
AYMELSSLRSEDTAVYYCARKLLNTVVVTDYAMDYWGQG
TTVTVSS
J647M2S1-65VH
918
EVQLVQSGAEVKKPGASVKVSCKASGYTFTNYGMNWVRQ
APGQGLEWMGWINTYTGEPTYADDFKGRFTFTLDTSTST
AYMELSSLRSEDTAVYYCARKFLTPVVVTDCAMDYWGQG
TTVTVSS
J647M2S1-66VH
919
EVQLVQSGAEVKKPGASVKVSCKASGYTFTNYGINWVRQ
APGQGLEWMGWINTYTGEPRYAQGLTGRVTMTTDTSTST
AYMELSSLRSEDTAVYFCARKFLTTVVVTDYAMDYWGQG
TTVTVSS
J647M2S1-67VH
920
EVQLVQSGAEVKKPGASVKVSCKASGYTFRDYGINWVRQ
APGQGLEWMGWINTYTGLPTYAQGFTGRVTMTTDTSTST
AYMELSSLRSEDTAVYFCARKFLTTVVVTDYAMDYWGQG
TTVTVSS
J647M2S1-69VH
921
EVQLVQSGAEVKKPGASVKVSCKASGYTFTNYGMNWVRQ
APGQGLEWMGWINTYTGEPTYADDFKGRFTFTLDTSTST
AYMELSSLRSEDTAVYFCARKFWTTIVVTDYAMDYWGQG
TTVTVSS
J647M2S1-6VH
922
EVQLVQSGAEVKKPGASVKVSCKASGYTFTNYGMNWVRQ
APGQGLEWMGWINTYTGEPTYADDFKGRFTFTLDTSTST
AYMELSSLRSEDTAVYFCARKLLTTVSATDNAMDYWGQG
TTVTVSS
J647M2S1-70VH
923
EVQLVQSGAEVKKPGASVKVSCKASGYTFTNYGMNWVRQ
APGQGLEWMGWINTYTGEPTYADDFKGRFTFTLDTSTST
AYMELSSLRSEDTAVYYCARKFLNTVVVTDYAMDYWGQG
TTVTVSS
J647M2S1-72VH
924
EVQLVQSGAEVKKPGASVKVSCKASGYTFTDYGINWVRQ
APGQGLEWMGWINTYNGEPSYAQGLTGRVTMTTDTSTST
AYMELSSLRSEDTAVYFCARKFLTTVVVTDYAMDYWGQG
TTVTVSS
J647M2S1-75VH
925
EVQLVQSGAEVKKPGASVKVSCKASGYTFATYGIAWVRQ
APGQGLEWMGWINTYSGVPKYAQGLTGRVTMTTDTSTST
AYMELSSLRSEDTAVYFCARKFLTTVVVTDYAMDYWGQG
TTVTVSS
J647M2S1-76VH
926
EVQLVQSGAEVKKPGASVKVSCKASGYTFTNYGMNWVRQ
APGQGLEWMGWINTYTGEPTYADDFKGRFTFTLDTSTST
AYMELSSLRSEDTAVYFCARKFRTTAVPTDNAMDYWGQG
TTVTVSS
J647M2S1-77VH
927
EVQLVQSGAEVKKPGASVKVSCKASGYTFTNYGMNWVRQ
APGQGLEWMGWINTYTGEPTYADDFKGRFTFTLDTSTST
AYMELSSLRSEDTAVYYCARKFLTTVVNTDSAMDYWGQG
TTVTVSS
J647M2S1-78VH
928
EVQLVQSGAEVKKPGASVKVSCKASGYTFTNYGMNWVRQ
APGQGLEWMGWINTYTGEPTYADDFKGRFTFTLDTSTST
AYMELSSLRSEDTAVYFCARKFLTTVVVTDYAMDYWGRG
TTVTVSS
J647M2S1-79VH
929
EVQLVQSGAEVKKPGASVKVSCKASGYTFTNYGMNWVRQ
APGQGLEWMGWINTYTGEPTYADDFKGRFTFTLDTSTST
AYMELSSLRSEDTAVYYCARKLLKTRVVTDYAMDYWGQG
TTVTVSS
J647M2S1-7VH
930
EVQLVQSGAEVKKPGASVKVSCKASGYTFTNYGMNWVRQ
APGQGLEWMGWINTYTGEPTYADDFKGRFTFTLDTSTST
AYMELSSLRSEDTAVYFCARKFLTTVVVTDYAMDYWGQG
TTVTVSS
J647M2S1-80VH
931
EVQLVQSGAEVKKPGASVKVSCKASGYTFTNYGMNWVRQ
APGQGLEWMGWINTYTGEPTYADDFKGRFTFTLDTSTST
AYMELSSLRSEDTAVYYCARKLLTTVVATDYAMDYWGQG
TTVTVSS
J647M2S1-84VH
932
EVQLVQSGAEVKKPGASVKVSCKASGYTFTNYGINWVRQ
APGQGLEWMGWINTYTGEPTYAQGFTGRVTMTTDTSTST
AYMELSSLRSEDTAVYFCARKFLTTVVVTDYAMDYWGQG
TTVTVSS
J647M2S1-85VH
933
EVQLVQSGAEVKKPGASVKVSCKASGYTFTNYGINWVRQ
APGQGLEWMGWINTYTGQPTYAQGFTGRVTMTTDTSTST
AYMELSSLRSEDTAVYFCARKFLTTVVVTDYAMDYWGQG
TTVTVSS
J647M2S1-87VH
934
EVQLVQSGAEVKKPGASVKVSCKASGYTFTNYGMNWVRQ
APGQGLEWMGWINTYTGEPTYADDFKGRFTFTLDTSTST
AYMELSSLRSEDTAVYYCARKFFPTMVVTDYAMDYWGQG
TTVTVSS
J647M2S1-88VH
935
EVQLVQSGAEVKKPGASVKVSCKASGYTFTNYGMNWVRQ
APGQGLEWMGWINTYTGEPTYADDFKGRFTFTLDTSTST
AYMELSSLRSEDTAVYYCARKFVTTMVVTDYAMDYWGQG
TTVTVSS
J647M2S1-8VH
936
EVQLVQSGAEVKKPGASVKVSCKASGYTFTNYGMNWVRQ
APGQGLEWMGWINTYTGEPTYAQGLTGRVTMTTDTSTST
AYMELSSLRSEDTAVYFCARKFLTTVVVTDYAMDYWGQG
TTVTVSS
J647M2S1-92VH
937
EVQLVQSGAEVKKPGASVKVSCKASGYTFTNYGMNWVRQ
APGQGLEWMGWINTYTGEPTYADDFKGRFTFTLDTSTST
AYMELSSLRSEDTAVYFCARKLLTTIVATDNAMDYWGQG
TTVTVSS
J647M2S1-93VH
938
EVQLVQSGAEVKKPGASVKVSCKASGYTFTNYGMNWVRQ
APGQGLEWMGWINTYTGEPTYADDFKGRFTFTLDTSTST
AYMELSSLRSEDTAVYYCARKLMSTVVETDNAMDYWGQG
TTVTVSS
J647M2S1-94VH
939
EVQLVQSGAEVKKPGASVKVSCKASGYTFTNYGMNWVRQ
APGQGLEWMGWINTYTGEPTYADDFKGRFTFTLDTSTST
AYMELSSLRSEDTAVYYCARKLLFTVVQTDYAMDYWGQG
TTVTVSS
J647M2S1-96VH
940
EVQLVQSGAEVKKPGASVKVSCKASGYTFTNYGMNWVRQ
APGQGLEWMGWINTYTGEPTYADDFKGRFTFTLDTSTST
AYMELSSLRSEDTAVYYCARKLLNTVVDTDYAMDYWGQG
TTVTVSS
J662M2S3-14VH
941
EVQLVQSGAEVKKPGASVKVSCKASGYTFSNYGIIWVRQ
APGQGLEWMGWINTYTGEPHYAQKLQGRVTMTTDTSTST
AYMELSSLRSEDTAVYFCARKLFTVEDVTDCAMDYWGQG
TTVTVSS
J662M2S3-18VH
942
EVQLVQSGAEVKKPGASVKVSCKASGYTFDNYGMNWVRQ
APGQGLEWMGWINTYNGKPTYAQKFQGRVTMTTDTSTST
AYMELSSLRSEDTAVYFCARKLFLVEAVTDYAMDYWGQG
TTVTVSS
J662M2S3-28VH
943
EVQLVQSGAEVKKPGASVKVSCKASGYTFRNYGIIWVRQ
APGQGLEWMGWINTYTGKPTYAQKFQGRVTMTTDTSTST
AYMELSSLRSEDTAVYFCARKLFTTVDVTDNAMDYWGQG
TTVTVSS
J662M2S3-29VH
944
EVQLVQSGAEVKKPGASVKVSCKASGYTFNNYGIIWVRQ
APGQGLEWMGWINTYTGVPTYAQKFQGRVTMTTDTSTST
AYMELSSLRSEDTAVYYCARKLFNTVDVTDNAMDYWGQG
TTVTVSS
J662M2S3-30VH
945
EVQLVQSGAEVKKPGASVKVSCKASGYTFNNYGIIWVRQ
APGQGLEWMGWINTYTGEPHYAQKFQGRVTMTTDTSTST
AYMELSSLRSEDTAVYYCARKLFKTMAVTDAAMDYWGQG
TTVTVSS
J662M2S3-34VH
946
EVQLVQSGAEVKKPGASVKVSCKASGYTFNNYGINWVRQ
APGQGLEWMGWINTYTGKPTYAQKFQGRVTMTTDTSTST
AYMELSSLRSEDTAVYFCARKFRNTVAVTDYAMDYWGQG
TTVTVSS
J662M2S3-3VH
947
EVQLVQSGAEVKKPGASVKVSCKASGYTFNNYGIIWVRQ
APGQGLEWMGWINTYTGKPTYAQKFQGRVTMTTDTSTST
AYMELSSLRSEDTAVYYCARKLFNTVAVTDNAMDYWGQG
TTVTVSS
J662M2S3-41VH
948
EVQLVQSGAEVKKPGASVKVSCKASGYTFTNYGINWVRQ
APGQGLEWMGWINTYTGKPTYAQKFQGRVTMTTDTSTST
AYMELSSLRSEDTAVYYCARKLFFTEDVTDYAMDYWGQG
TTVTVSS
J662M2S3-45VH
949
EVQLVQSGAEVKKPGASVKVSCKASGYTFNNYGINWVRQ
APGQGLEWMGWINTYTGKPTYAQKFQGRVTMTTDTSTST
AYMELSSLRSEDTAVYFCARKFFTPVVVTDNAMDYWGQG
TTVTVSS
J662M2S3-55VH
950
EVQLVQSGAEVKKPGASVKVSCKASGYTFRNYGITWVRQ
APGQGLEWMGWINTYTGKPTYAQKFQGRVTMTTDTSTST
AYMELSSLRSEDTAVYFCARKLFTTMDVTDNAMDYWGQG
TTVTVSS
J662M2S3-5VH
951
EVQLVQSGAEVKKPGASVKVSCKASGYTFANYGIIWVRQ
APGQGLEWMGWINTYTGKPTYAQKFQGRVTMTTDTSTST
AYMELSSLRSEDTAVYYCARKLFTTMDVTDNAMDYWGQG
TTVTVSS
J662M2S3-65VH
952
EVQLVQSGAEVKKPGASVKVSCKASGYTFNNYGIIWVRQ
APGQGLEWMGWINTYTGKPTYAQKLQGRVTMTTDTSTST
AYMELSSLRSEDTAVYFCARKLFNTVDVTDNAMDYWGQG
TTVTVSS
J662M2S3-78VH
953
EVQLVQSGAEVKKPGASVKVSCKASGYTFSNYGIIWVRQ
APGQGLEWMGWINTYTGKPSYAQKFQGRVTMTTDTSTST
AYMELSSLRSEDTAVYYCARKLFNTVDVTDNAMDYWGQG
TTVTVSS
J662M2S3-84VH
954
EVQLVQSGAEVKKPGASVKVSCKASGYTFSNYGINWVRQ
APGQGLEWMGWINTYTGQPSYAQKFQGRVTMTTDTSTST
AYMELSSLRSEDTAVYYCARKLFKTEAVTDYAMDYWGQG
TTVTVSS
J662M2S3-87VH
955
EVQLVQSGAEVKKPGASVKVSCKASGYTFNNYGIIWVRQ
APGQGLEWMGWINTYSGKPTYAQKFQGRVTMTTDTSTST
AYMELSSLRSEDTAVYFCARKLFTTMDVTDNAMDYWGQG
TTVTVSS
J662M2S3-96VH
956
EVQLVQSGAEVKKPGASVKVSCKASGYTFNNYGIIWVRQ
APGQGLEWMGWINTYTGKPTYAQKFQGRVTMTTDTSTST
AYMELSSLRSEDTAVYYCARKFFTTMAVTDNAMDYWGQG
TTVTVSS
Table 28 provides a list of amino acid sequences of VL regions of affinity matured fully human TNF antibodies derived from hMAK199 Amino acid residues of individual CDRs of each VL sequence are indicated in bold.
TABLE 28
List of amino acid sequences of affinity matured hMAK199
VL variants
Clone
SEQ ID NO:
VL
J644M2S1-11Vk
957
DIQMTQSPSSLSASVGDRVTITCRASQDISNYLNWYQQK
PGKAPKLLIYYTSRLQSGVPSRFSGSGSGTDYTLTISSL
QPEDFATYYCQQGNTLPPTFGQGTKLEIK
J644M2S1-73Vk
958
DIQMTQSPSSLSASVGDRVTITCRASQDISNYLNWYQQK
PGKAPKLLIYYASRLQSGVPSRFSGSGSGTDYTLTISSL
QPEDFATYFCQQGNTLPPTFGQGTKLEIK
J647M2-11Vk
959
DIQMTQSPSSLSASVGDRVTITCRASQDISNYLNWYQQK
PGKTVKLLIYYTSRLQSGVPSRFSGSGSGTDYTLTISSL
QPEDFATYFCQQGNTLPPTFGQGTKLEIK
J647M2S1-10Vk
960
DIQMTQSPSSLSASVGDRVTITCRASQDIWNYLNWYQQK
PGKAPKLLIYYTSRLQSGVPSRFSGSGSGTDFTLTISSL
QPEDFATYFCQQGNRYPPTFGQGTKLEIK
J647M2S1-16Vk
961
DIQMTQSPSSLSASVGDRVTITCRASQDICTYLNWYQQK
PGKAPKLLIYYTSRLQSGVPSRFSGSGSGTDFTLTISSL
QPEDFATYYCQQGNSPPPTFGQGTKLEIK
J647M2S1-1Vk
962
DIQMTQSPSSLSASVGDRVTITCRASQAIGNYLNWYQQK
PGKAPKLLIYYASRLQSGVPSRFSGSGSGTDFTLTISSL
QPEDFATYFCQQGNTLPPTFGQGTKLEIK
J647M2S1-20Vk
963
DIQMTQSPSSLSASVGDRVTITCRASQDISNYLNWYQQK
PGKAPKLLIYYTSRLQSGVPSRFSGSGSGTDYTLTISSL
QPEDFATYFCQQGNTRPPTFGQGTKLEIK
J647M2S1-24Vk
964
DIQMTQSPSSLSASVGDRVTITCRASQDISNYLNWYQQK
PGKAPKLLIYYTSLLQSGVPSRFSGSGSGTDYTLTISSL
QPEDFATYYCQQGNTGPPTFGQGTKLEIK
J647M2S1-25Vk
965
DIQMTQSPSSLSASVGDRVTITCRASQDIYNYLNWYQQK
PGKAPKLLIYYTSRLQSGVPSRFSGSGSGTDYTLTISSL
QPEDFATYFCQQGNTLPPTFGQGTKLEIK
J647M2S1-29Vk
966
DIQMTQSPSSLSASVGDRVTITCRASQDISHYLNWYQQK
PGKAPKLLIYYTSRLQSGVPSRFSGSGSGTDYTLTISSL
QPEDFATYFCQQGNTLPATFGQGTKLEIK
J647M2S1-2Vk
967
DIQMTQSPSSLSASVGDRVTITCRASQDISSYLNWYQQK
PGKTVKLLIYYASRLQSGVPSRFSGSGSGTDFTLTISSL
QPEDFATYYCQQGNTPPPTFGQGTKLEIK
J647M2S1-34Vk
968
DIQMTQSPSSLSASVGDRVTITCRASQDISSYLNWYQQK
PGKAPKLLIYYASRLQSGVPSRFSGSGSGTDFTLTISSL
QPEDFATYYCQQGNTLPPTFGQGTKLEIK
J647M2S1-37Vk
969
DIQMTQSPSSLSASVGDRVTITCRASQEISNYLNWYQQK
PGKAPKLLIYYTSRLQSGVPSRFSGSGSGTDYTLTISSL
QPEDFATYFCQQGNTMPTTFGQGTKLEIK
J647M2S1-38Vk
970
DIQMTQSPSSLSASVGDRVTITCRASQDISNYLNWYQQK
PGKAPKLLIYFASRLQSGVPSRFSGSGSGTDYTLTISSL
QPEDFATYFCQQGNTPPTTFGQGTKLEIK
J647M2S1-3Vk
971
DIQMTQSPSSLSASVGDRVTITCRASQDISNYLNWYQQK
PGKAPKLLIYYTSRLQSGVPSRFSGSGSGTDYTLTISSL
QPEDFATYFCQQGNTLPSTFGQGTKLEIK
J647M2S1-42Vk
972
DIQMTQSPSSLSASVGDRVTITCRASQVISNTLNWYQQK
PGKAPKLLIYYTSRLQSGVPSRFSGSGSGTDYTLTISSL
QPEDFATYFCQQGNALPPTFGQGTKLEIK
J647M2S1-44Vk
973
DIQMTQSPSSLSASVGDRVTITCRASQDISTYLNWYQQK
PGKAPKLLIYYTSRLQSGVPSRFSGSGSGTDFTLTISSL
QPEDFATYFCQQGNTPPPTFGQGTKLEIK
J647M2S1-46Vk
974
DIQMTQSPSSLSASVGDRVTITCRASQDISQYLNWYQQK
PGKAPKLLIYYTSRLQSGVPSRFSGSGSGTDYTLTISSL
QPEDFATYYCQQGNTLPPTFGQGTKLEIK
J647M2S1-50Vk
975
DIQMTQSPSSLSASVGDRVTITCRASQDITNYLNWYQQK
PGKAPKLLIYYTSRLQSGVPSRFSGSGSGTDFTLTISSL
QPEDFATYYCQQGNTAPPTFGQGTKLEIK
J647M2S1-52Vk
976
DIQMTQSPSSLSASVGDRVTITCRASQGISNYLNWYQQK
PGKAPKLLIYYTSRLQSGVPSRFSGSGSGTDYTLTISSL
QPEDFATYFCQQGNTMPPTFGQGTKLEIK
J647M2S1-56Vk
977
DIQMTQSPSSLSASVGDRVTITCRASQDISNYLNWYQQK
PGKAPKLLIYYTSRLQSGVPSRFSGSGSGTDFTLTISSL
QPEDFATYFCQQGNTLPPTFGQGTKLEIK
J647M2S1-59Vk
978
DIQMTQSPSSLSASVGDRVTITCRASQDISKYLNWYQQK
PGKAPKLLIYYTSRLQSGVPSRFSGSGSGTDFTLTISSL
QPEDFATYYCQQGNTRPPTFGQGTKLEIK
J647M2S1-71Vk
979
DIQMTQSPSSLSASVGDRVTITCRASQDISNYLNWYQQK
PGKAPKLLIYYTSLLQSGVPSRFSGSGSGTDFTLTISSL
QPEDFATYYCQQGNTQPPTFGQGTKLEIK
J647M2S1-74Vk
980
DIQMTQSPSSLSASVGDRVTITCRASQDISNYLNWYQQK
PGKAPKLLIYYTSRLQSGVPSRFSGSGSGTDFTLTISSL
QPEDFATYYCQQGNSQPPTFGQGTKLEIK
J647M2S1-78Vk
981
DIQMTQSPSSLSASVGDRVTITCRASQDISKYLNWYQQK
PGKAPKLLIYNASRLQSGVPSRFSGSGSGTDFTLTISSL
QPEDFATYFCQQGNTLPPTFGQGTKLEIK
J647M2S1-7Vk
982
DIQMTQSPSSLSASVGDRVTITCRASQDISNYLNWYQQK
PGKAPKLLIYYTSLLQSGVPSRFSGSGSGTDFTLTISSL
QPEDFATYYCQQGNIWPPTFGQGTKLEIK
J647M2S1-9Vk
983
DIQMTQSPSSLSASVGDRVTITCRASQDISHYLNWYQQK
PGKAPKLLIYYTSRLQSGVPSRFSGSGSGTDYTLTISSL
QPEDFATYFCQQGNTLPPTFGQGTKLEIK
J652M2S1-10Vk
984
DIQMTQSPSSLSASVGDRVTITCRASQDISNYLNWYQQK
PGKAPKLLIYYTSRLQSGVPSRFSGSGSGTDYTLTISSL
QPEDFATYYCQQGNTFPPTFGQGTKLEIK
J652M2S1-13Vk
985
DIQMTQSPSSLSASVGDRVTITCRASQDISNYLNWYQQK
PGKAPKLLIYYTSRLQSGVPSRFSGSGSGTDYTLTISSL
QPEDFATYFCQQGNTQPPTFGQGTKLEIK
J652M2S1-14Vk
986
DIQMTQSPSSLSASVGDRVTITCRASQDISNVLNWYQQK
PGKAPKLLIYYASRLQSGVPSRFSGSGSGTDFTLTISSL
QPEDFATYYCQQGNTLPPTFGQGTKLEIK
J652M2S1-15Vk
987
DIQMTQSPSSLSASVGDRVTITCRASQDIYKYLNWYQQK
PGKAPKLLIYYTSRLQSGVPSRFSGSGSGTDFTLTISSL
QPEDFATYYCQQGNTMPPTFGQGTKLEIK
J652M2S1-17Vk
988
DIQMTQSPSSLSASVGDRVTITCRASQEIFSYLNWYQQK
PGKAPKLLIYYTSRLQSGVPSRFSGSGSGTDFTLTISSL
QPEDFATYYCQQGNMGPPTFGQGTKLEIK
J652M2S1-18Vk
989
DIQMTQSPSSLSASVGDRVTITCRASQDISNYLNWYQQK
PGKAPKLLIYYTSRLQSGVPSRFSGSGSGTDYTLTISSL
QPEDFATYYCQQGNTQPPTFGQGTKLEIK
J652M2S1-1Vk
990
DIQMTQSPSSLSASVGDRVTITCRASQDISSYLNWYQQK
PGKAPKLLIYYASRLQSGVPSRFSGSGSGTDFTLTISSL
QPEDFATYYCQQGNTWPPTFGQGTKLEIK
J652M2S1-22Vk
991
DIQMTQSPSSLSASVGDRVTITCRASQDISNYLNWYQQK
PGKAPKLLIYYTSRLQSGVPSRFSGSGSGTDFTLTISSL
QPEDFATYYCQQGNTRPPTFGQGTKLEIK
J652M2S1-23Vk
992
DIQMTQSPSSLSASVGDRVTITCRASQDISNFLNWYQQK
PGKAPKLLIYYASRLQSGVPSRFSGSGSGTDFTLTISSL
QPEDFATYFCQQGNTFPPTFGQGTKLEIK
J652M2S1-25Vk
993
DIQMTQSPSSLSASVGDRVTITCRASQDISNFLNWYQQK
PGKAPKLLIYYTSRLQSGVPSRFSGSGSGTDYTLTISSL
QPEDFATYFCQQGNTRPPTFGQGTKLEIK
J652M2S1-26Vk
994
DIQMTQSPSSLSASVGDRVTITCRASQDINNYLNWYQQK
PGKAPKLLIYYTSRLQSGVPSRFSGSGSGTDFTLTISSL
QPEDFATYFCQQGNTQPPTFGQGTKLEIK
J652M2S1-27Vk
995
DIQMTQSPSSLSASVGDRVTITCRASQDISNYLNWYQQK
PGKAPKLLIYYASGLQSGVPSRFSGSGSGTDYTLTISSL
QPEDFATYFCQQGNTWPPTFGQGTKLEIK
J652M2S1-28Vk
996
DIQMTQSPSSLSASVGDRVTITCRASQDISRYLNWYQQK
PGKAPKLLIYYTSRLQSGVPSRFSGSGSGTDFTLTISSL
QPEDFATYYCQQGNTQPPTFGQGTKLEIK
J652M2S1-29Vk
997
DIQMTQSPSSLSASVGDRVTITCRASQDIATYLNWYQQK
PGKAPKLLIYYTSRLQSGVPSRFSGSGSGTDFTLTISSL
QPEDFATYFCQQGNTMPPTFGQGTKLEIK
J652M2S1-31Vk
998
DIQMTQSPSSLSASVGDRVTITCRASQDISNYLNWYQQK
PGKAPKLLIYYTSRLQSGVPSRFSGSGSGTDYTLTISSL
QPEDFATYFCQQGNTFPPTFGQGTKLEIK
J652M2S1-33Vk
999
DIQMTQSPSSLSASVGDRVTITCRASQRIGNYLNWYQQK
PGKTVKLLIYYASRLQSGVPSRFSGSGSGTDFTLTISSL
QPEDFATYFCQQGNTLPPTFGQGTKLEIK
J652M2S1-34Vk
1000
DIQMTQSPSSLSASVGDRVTITCRASQEISNYLNWYQQK
PGKAPKLLIYYTSRLQSGVPSRFSGSGSGTDYTLTISSL
QPEDFATYYCQQGNSQPPTFGQGTKLEIK
J652M2S1-35Vk
1001
DIQMTQSPSSLSASVGDRVTITCRASQDIANYLNWYQQK
PGKAPKLLIYYTSRLQSGVPSRFSGSGSGTDFTLTISSL
QPEDFATYYCQQGNTLPPTFGQGTKLEIK
J652M2S1-37Vk
1002
DIQMTQSPSSLSASVGDRVTITCRASQDISNYLNWYQQK
PGKAPKLLIYYASRLQSGVPSRFSGSGSGTDYTLTISSL
QPEDFATYFCQQGNTFPPTFGQGTKLEIK
J652M2S1-38Vk
1003
DIQMTQSPSSLSASVGDRVTITCRASQDISNYLNWYQQK
PGKAPKLLIYYTSRLQSGVPSRFSGSGSGTDFTLTISSL
QPEDFATYYCQQGNTQPPTFGQGTKLEIK
J652M2S1-3Vk
1004
DIQMTQSPSSLSASVGDRVTITCRASQDISNYLNWYQQK
PGKAPKLLIYYTSRLQSGVPSRFSGSGSGTDYTLTISSL
QPEDFATYFCQQGNTPPPTFGQGTKLEIK
J652M2S1-40Vk
1005
DIQMTQSPSSLSASVGDRVTITCRASQDISNFLNWYQQK
PGKAPKLLIYYTSRLQSGVPSRFSGSGSGTDYTLTISSL
QPEDFATYYCQQGNTLPPTFGQGTKLEIK
J652M2S1-41Vk
1006
DIQMTQSPSSLSASVGDRVTITCRASQDIGNFLNWYQQK
PGKAPKLLIYYTSRLQSGVPSRFSGSGSGTDYTLTISSL
QPEDFATYFCQQGNTRPPTFGQGTKLEIK
J652M2S1-42Vk
1007
DIQMTQSPSSLSASVGDRVTITCRASQDITNYLNWYQQK
PGKAPKLLIYYTSRLQSGVPSRFSGSGSGTDYTLTISSL
QPEDFATYYCQQGNTPPPTFGQGTKLEIK
J652M2S1-45Vk
1008
DIQMTQSPSSLSASVGDRVTITCRASQDISDYLNWYQQK
PGKAPKLLIYYTSRLQSGVPSRFSGSGSGTDYTLTISSL
QPEDFATYFCQQGNMWPPTFGQGTKLEIK
J652M2S1-47Vk
1009
DIQMTQSPSSLSASVGDRVTITCRASQDISSYLNWYQQK
PGKAPKLLIYYASRLQSGVPSRFSGSGSGTDFTLTISSL
QPEDFATYFCQQGNTMPPTFGQGTKLEIK
J652M2S1-48Vk
1010
DIQMTQSPSSLSASVGDRVTITCRASQDISHYLNWYQQK
PGKAPKLLIYYTSRLQSGVPSRFSGSGSGTDFTLTISSL
QPEDFATYYCQQGNTLPPTFGQGTKLEIK
J652M2S1-49Vk
1011
DIQMTQSPSSLSASVGDRVTITCRASQDISNYLNWYQQK
PGKAPKLLIYYASRLQSGVPSRFSGSGSGTDFTLTISSL
QPEDFATYFCQQGNTMPPTFGQGTKLEIK
J652M2S1-51Vk
1012
DIQMTQSPSSLSASVGDRVTITCRASQDISQYLNWYQQK
PGKAPKLLIYYTSRLQSGVPSRFSGSGSGTDFTLTISSL
QPEDFATYFCQQGNTRPPTFGQGTKLEIK
J652M2S1-52Vk
1013
DIQMTQSPSSLSASVGDRVTITCRASQDISNYLNWYQQK
PGKAPKLLIYYTSRLQSGVPSRFSGSGSGTDYTLTISSL
QPEDFATYFCQQGNMRPPTFGQGTKLEIK
J652M2S1-53Vk
1014
DIQMTQSPSSLSASVGDRVTITCRASQDISTYLNWYQQK
PGKAPKLLIYYASRLQSGVPSRFSGSGSGTDFTLTISSL
QPEDFATYFCQQGNTLPPTFGQGTKLEIK
J652M2S1-55Vk
1015
DIQMTQSPSSLSASVGDRVTITCRASQDISNYLNWYQQK
PGKAPKLLIYYTSRLQSGVPSRFSGSGSGTDYTLTISSL
QPEDFATYFCQQGNTGPPTFGQGTKLEIK
J652M2S1-56Vk
1016
DIQMTQSPSSLSASVGDRVTITCRASQNINNYLNWYQQK
PGKAPKLLIYYTSRLQSGVPSRFSGSGSGTDYTLTISSL
QPEDFATYFCQQGNTMPPTFGQGTKLEIK
J652M2S1-57Vk
1017
DIQMTQSPSSLSASVGDRVTITCRASQDISKYLNWYQQK
PGKAPKLLIYYTSRLQSGVPSRFSGSGSGTDYTLTISSL
QPEDFATYYCQQGNTPPPTFGQGTKLEIK
J652M2S1-61Vk
1018
DIQMTQSPSSLSASVGDRVTITCRASQDISNYLNWYQQK
PGKAPKLLIYYTSRLQSGVPSRFSGSGSGTDFTLTISSL
QPEDFATYYCQQGNTVPPTFGQGTKLEIK
J652M2S1-62Vk
1019
DIQMTQSPSSLSASVGDRVTITCRASQDISNYLNWYQQK
PGKAPKLLIYYTSKLQSGVPSRFSGSGSGTDYTLTISSL
QPEDFATYFCQQGNIFPPTFGQGTKLEIK
J652M2S1-64Vk
1020
DIQMTQSPSSLSASVGDRVTITCRASQGIYNYLNWYQQK
PGKAPKLLIYYTSRLQSGVPSRFSGSGSGTDYTLTISSL
QPEDFATYFCQQGNTLPPTFGQGTKLEIK
J652M2S1-67Vk
1021
DIQMTQSPSSLSASVGDRVTITCRASQDISSYLNWYQQK
PGKAPKLLIYYTSRLQSGVPSRFSGSGSGTDFTLTISSL
QPEDFATYYCQQGNTLPPTFGQGTKLEIK
J652M2S1-69Vk
1022
DIQMTQSPSSLSASVGDRVTITCRASQEISNYLNWYQQK
PGKAPKLLIYYTSRLQSGVPSRFSGSGSGTDYTLTISSL
QPEDFATYFCQQGNTGPPTFGQGTKLEIK
J652M2S1-6Vk
1023
DIQMTQSPSSLSASVGDRVTITCRASQDISNYLNWYQQK
PGKAPKLLIYYTSRLQSGVPSRFSGSGSGTDFTLTISSL
QPEDFATYYCQQGNTPPPTFGQGTKLEIK
J652M2S1-71Vk
1024
DIQMTQSPSSLSASVGDRVTITCRASQDISDYLNWYQQK
PGKAPKLLIYYTSRLQSGVPSRFSGSGSGTDYTLTISSL
QPEDFATYYCQQGNTWPPTFGQGTKLEIK
J652M2S1-73Vk
1025
DIQMTQSPSSLSASVGDRVTITCRASQDIWKYLNWYQQK
PGKAPKLLIYYASRLQSGVPSRFSGSGSGTDFTLTISSL
QPEDFATYYCQQGNTLPPTFGQGTKLEIK
J652M2S1-75Vk
1026
DIQMTQSPSSLSASVGDRVTITCRASQDISTYLNWYQQK
PGKAPKLLIYYTSRLQSGVPSRFSGSGSGTDYTLTISSL
QPEDFATYYCQQGNTWPPTFGQGTKLEIK
J652M2S1-77Vk
1027
DIQMTQSPSSLSASVGDRVTITCRASQDISNYLNWYQQK
PGKAPKLLIYYTSRLQSGVPSRFSGSGSGTDYTLTISSL
QPEDFATYYCQQGNTPPPTFGQGTKLEIK
J652M2S1-78Vk
1028
DIQMTQSPSSLSASVGDRVTITCRASQDISNYLNWYQQK
PGKAPKLLIYYTSRLQSGVPSRFSGSGSGTDFTLTISSL
QPEDFATYFCQQGNAPPPTFGQGTKLEIK
J652M2S1-79Vk
1029
DIQMTQSPSSLSASVGDRVTITCRASQDIYKFLNWYQQK
PGKAPKLLIYYTSRLQSGVPSRFSGSGSGTDYTLTISSL
QPEDFATYFCQQGNTLPPTFGQGTKLEIK
J652M2S1-80Vk
1030
DIQMTQSPSSLSASVGDRVTITCRASQDIFNYLNWYQQK
PGKAPKLLIYYTSRLQSGVPSRFSGSGSGTDYTLTISSL
QPEDFATYFCQQGNTLPPTFGQGTKLEIK
J652M2S1-82Vk
1031
DIQMTQSPSSLSASVGDRVTITCRASQDISNTLNWYQQK
PGKAPKLLIYYASRLQSGVPSRFSGSGSGTDFTLTISSL
QPEDFATYFCQQGNTLPPTFGQGTKLEIK
J652M2S1-84Vk
1032
DIQMTQSPSSLSASVGDRVTITCRASQHISNYLNWYQQK
PGKAPKLLIYYTSRLQSGVPSRFSGSGSGTDYTLTISSL
QPEDFATYFCQQGNTQPPTFGQGTKLEIK
J652M2S1-86Vk
1033
DIQMTQSPSSLSASVGDRVTITCRASQDISNYLNWYQQK
PGKAPKLLIYYTSRLQSGVPSRFSGSGSGTDYTLTISSL
QPEDFATYFCQQGNMPPPTFGQGTKLEIK
J652M2S1-87Vk
1034
DIQMTQSPSSLSASVGDRVTITCRASQDITNYLNWYQQK
PGKAPKLLIYYTSRLQSGVPSRFSGSGSGTDFTLTISSL
QPEDFATYFCQQGNTVPPTFGQGTKLEIK
J652M2S1-8Vk
1035
DIQMTQSPSSLSASVGDRVTITCRASQDISNYLNWYQQK
PGKAPKLLIYFTSRLQSGVPSRFSGSGSGTDFTLTISSL
QPEDFATYYCQQGNTQPPTFGQGTKLEIK
J652M2S1-90Vk
1036
DIQMTQSPSSLSASVGDRVTITCRASQDISKFLNWYQQK
PGKAPKLLIYYTSRLQSGVPSRFSGSGSGTDYTLTISSL
QPEDFATYYCQQGNTRPPTFGQGTKLEIK
J652M2S1-91Vk
1037
DIQMTQSPSSLSASVGDRVTITCRASQDISNYLNWYQQK
PGKAPKLLIYYTSRLQSGVPSRFSGSGSGTDFTLTISSL
QPEDFATYFCQQGNTFPPTFGQGTKLEIK
J652M2S1-92Vk
1038
DIQMTQSPSSLSASVGDRVTITCRASQDIYNVLNWYQQK
PGKAPKLLIYYASRLQSGVPSRFSGSGSGTDFTLTISSL
QPEDFATYFCQQGITLPPTFGQGTKLEIK
J652M2S1-93Vk
1039
DIQMTQSPSSLSASVGDRVTITCRASQHISNYLNWYQQK
PGKAPKLLIYYTSRLQSGVPSRFSGSGSGTDYTLTISSL
QPEDFATYFCQQGNTWPPTFGQGTKLEIK
J652M2S1-95Vk
1040
DIQMTQSPSSLSASVGDRVTITCRASQDISNYLNWYQQK
PGKAPKLLIYYTSRLQSGVPSRFSGSGSGTDYTLTISSL
QPEDFATYFCQQGNTQPSTFGQGTKLEIK
J652M2S1-9Vk
1041
DIQMTQSPSSLSASVGDRVTITCRASQDISNYLNWYQQK
PGKAPKLLIYYTSRLQSGVPSRFSGSGSGTDFTLTISSL
QPEDFATYFCQQGNTQPPTFGQGTKLEIK
J662M2S3-13Vk
1042
DIQMTQSPSSLSASVGDRVTITCRASQDISNYLNWYQQK
PGKAPKLLIYYTSRLQSGVPSRFSGSGSGTDYTLTISSL
QPEDFATYFCQQGNSWPPTFGQGTKLEIK
J662M2S3-15Vk
1043
DIQMTQSPSSLSASVGDRVTITCRASQDIYNYLNWYQQK
PGKAPKLLIYYTSRLQSGVPSRFSGSGSGTDYTLTISSL
QPEDFATYFCQQGNTQPPTFGQGTKLEIK
J662M2S3-21Vk
1044
DIQMTQSPSSLSASVGDRVTITCRASQDISNYLNWYQQK
PGKAPKLLIYYTSRLQSGVPSRFSGSGSGTDYTLTISSL
QPEDFATYFCQQGNTWPPTFGQGTKLEIK
J662M2S3-22Vk
1045
DIQMTQSPSSLSASVGDRVTITCRASQDISQYLNWYQQK
PGKAPKLLIYYTSRLQSGVPSRFSGSGSGTDFTLTISSL
QPEDFATYFCQQGNTWPPTFGQGTKLEIK
J662M2S3-34Vk
1046
DIQMTQSPSSLSASVGDRVTITCRASQDIYDVLNWYQQK
PGKAPKLLIYYASRLQSGVPSRFSGSGSGTDFTLTISSL
QPEDFATYYCQQGITLPPTFGQGTKLEIK
J662M2S3-3Vk
1047
DIQMTQSPSSLSASVGDRVTITCRASQDIENYLNWYQQK
PGKAPKLLIYYTSRLQSGVPSRFSGSGSGTDFTLTISSL
QPEDFATYFCQQGNTQPPTFGQGTKLEIK
J662M2S3-41Vk
1048
DIQMTQSPSSLSASVGDRVTITCRASQNIENFLNWYQQK
PGKAPKLLIYYTSRLQSGVPSRFSGSGSGTDFTLTISSL
QPEDFATYFCQQGNTWPPTFGQGTKLEIK
J662M2S3-56Vk
1049
DIQMTQSPSSLSASVGDRVTITCRASQDIYNYLNWYQQK
PGKAPKLLIYYTSRLQSGVPSRFSGSGSGTDYTLTISSL
QPEDFATYFCQQGNTPPPTFGQGTKLEIK
J662M2S3-64Vk
1050
DIQMTQSPSSLSASVGDRVTITCRASQDIASYLNWYQQK
PGKAPKLLIYYTSRLQSGVPSRFSGSGSGTDFTLTISSL
QPEDFATYFCQQGNTQPPTFGQGTKLEIK
J662M2S3-78Vk
1051
DIQMTQSPSSLSASVGDRVTITCRASQDISNYLNWYQQK
PGKVPKLLIYYTSRLQSGVPSRFSGSGSGTDYTLTISSL
QPEDFATYFCQQGNTQPPTFGQGTKLEIK
J662M2S3-84Vk
1052
DIQMTQSPSSLSASVGDRVTITCRASQNIYNVLNWYQQK
PGKAPKLLIYYASRLQSGVPSRFSGSGSGTDFTLTISSL
QPEDFATYFCQQGNTMPPTFGQGTKLEIK
TABLE 29
Amino acid residues observed in affinity matured hMAK-199 antibodies
MAK199 Heavy chain variable region (SEQ ID NO: 1077)
MAK199VH.2a
1234567890123456789012345678901234567890123456789012a345678901
EIQLVQSGAEVKKPGASVKVSCKASGYTFTNYGMNWVRQAPGQGLEWMGWINTYTGEPTYAD
V ND II N K S Q
AH T S V H
ST Q Q N
RS S R M
DQ G L K
KK A S A
P V N R
Q I Q
M D D
G A
E
34567890123456789012abc345678901234567890abcdefg12345678901234
DFKGRFTFTLDTSTSTAYMELSSLRSEDTAVYFCARKFLTTVVVTDYAMDYWGQGTTVTVSS
GLT V M T Y RLFNPMDASENT
K Q NYMKVEAEM SR
IRSSAEMN CC
VSRARSD H
CWL IMG D
QP QII I
VF GPQ F
ND D P V
GM N L
CA L A
H
Mak199 Light chain variable region (SEQ ID NO: 1078)
Mak199Vk.1a
1234567890123456789012345678901234567890123456789012345678901
DIQMTQSPSSLSASVGDRVTITCRASQDISNYLNWYQQKPGKTVKLLIYYTSRLQSGVPSR
N YQV AP FA L
E ESF V N K
H AKT G
G TT
V WH
R GD
A NR
F
C
2345678901234567890123456789012345678901234567
FSGSGSGTDYTLTISSLQPEDFATYFCQQGNTLPPTFGQGTKLEIK
F Y ISW T
MQ S
IP A
AM
RR
F
G
V
Y
A
TABLE 30
Individual hMAK-199 VII sequences from converted clones
Protein
Sequence
region
SEQ ID NO:
123456789012345678901234567890
J662M2S3
1053
EVQLVQSGAEVKKPGASVKVSCKASGYTFA
#10 VH
NYGIIWVRQAPGQGLEWMGWINTYTGKPTY
AQKFQGRVTMTTDTSTSTAYMELSSLRSED
TAVYYCARKLFTTMDVTDNAMDYWGQGTTV
TVSS
J662M2S3#
CDR-H1
Residues 31-35
NYGII
10 VH
of SEQ ID
NO.: 1053
J662M2S3#
CDR-H2
Residues 50-66
WINTYTGKPTYAQKFQG
10 VH
of SEQ ID
NO.: 1053
J662M2S3#
CDR-H3
Residues 99-112
RASQDISQYLN
10 VH
of SEQ ID
NO.: 1053
J662M2S3#
1054
EVQLVQSGAEVKKPGASVKVSCKASGYTFN
13 VH
NYGIIWVRQAPGQGLEWMGWINTYTGKPTY
AQKLQGRVTMTTDTSTSTAYMELSSLRSED
TAVYFCARKLFNTVDVTDNAMDYWGQGTTV
TVSS
J662M2S3#
CDR-H1
Residues 31-35
NYGII
13 VH
of SEQ ID
NO.: 1054
J662M2S3#
CDR-H2
Residues 50-66
WINTYTGKPTYAQKLQG
13 VH
of SEQ ID
NO.: 1054
J662M2S3#
CDR-H3
Residues 99-112
KLFNTVDVTDNAMD
13 VH
of SEQ ID
NO.: 1054
J662M2S3#
1055
EVQLVQSGAEVKKPGASVKVSCKASGYTFN
15 VH
NYGIIWVRQAPGQGLEWMGWINTYTGVPTY
AQKFQGRVTMTTDTSTSTAYMELSSLRSED
TAVYYCARKLFNTVDVTDNAMDYWGQGTTV
TVSS
J662M2S3#
CDR-H1
Residues 31-35
NYGII
15 VH
of SEQ ID
NO.: 1055
J662M2S3#
CDR-H2
Residues 50-66
WINTYTGVPTYAQKFQG
15 VH
of SEQ ID
NO.: 1055
J662M2S3#
CDR-H3
Residues 99-112
KLFNTVDVTDNAMD
15 VH
of SEQ ID
NO.: 1055
J662M2S3#
1056
EVQLVQSGAEVKKPGASVKVSCKASGYTFN
16 VH
NYGIIWVRQAPGQGLEWMGWINTYTGKPTY
AQKFQGRVTMTTDTSTSTAYMELSSLRSED
TAVYYCARKLFNTVAVTDNAMDYWGQGTTV
TVSS
J662M2S3#
CDR-H1
Residues 31-35
NYGII
16 VH
of SEQ ID
NO.: 1056
J662M2S3#
CDR-H2
Residues 50-66
WINTYTGKPTYAQKFQG
16 VH
of SEQ ID
NO.: 1056
J662M2S3#
CDR-H3
Residues 99-112
KLFNTVAVTDNAMD
16 VH
of SEQ ID
NO.: 1056
J662M2S3#
1057
EVQLVQSGAEVKKPGASVKVSCKASGYTFR
21 VH
NYGIIWVRQAPGQGLEWMGWINTYTGKPTY
AQKFQGRVTMTTDTSTSTAYMELSSLRSED
TAVYFCARKLFTTVDVTDNAMDYWGQGTTV
TVSS
J662M2S3#
CDR-H1
Residues 31-35
NYGII
21 VH
of SEQ ID
NO.: 1057
J662M2S3#
CDR-H2
Residues 50-66
WINTYTGKPTYAQKFQG
21 VH
of SEQ ID
NO.: 1057
J662M2S3#
CDR-H3
Residues 99-112
KLFTTVDVTDNAMD
21 VH
of SEQ ID
NO.: 1057
J662M2S3#
1058
EVQLVQSGAEVKKPGASVKVSCKASGYTFN
34 VH
NYGINWVRQAPGQGLEWMGWINTYTGKPTY
AQKFQGRVTMTTDTSTSTAYMELSSLRSED
TAVYFCARKFRNTVAVTDYAMDYWGQGTTV
TVSS
J662M2S3#
CDR-H1
Residues 31-35
NYGIN
34 VH
of SEQ ID
NO.: 1058
J662M2S3#
CDR-H2
Residues 50-66
WINTYTGKPTYAQKFQG
34 VH
of SEQ ID
NO.: 1058
J662M2S3#
CDR-H3
Residues 99-112
KFRNTVAVTDYAMD
34 VH
of SEQ ID
NO.: 1058
J662M2S3#
1059
EVQLVQSGAEVKKPGASVKVSCKASGYTFR
36 VH
NYGITWVRQAPGQGLEWMGWINTYTGKPTY
AQKFQGRVTMTTDTSTSTAYMELSSLRSED
TAVYFCARKLFTTMDVTDNAMDYWGQGTTV
TVSS
J662M2S3#
CDR-H1
Residues 31-35
NYGIT
36 VH
of SEQ ID
NO.: 1059
J662M2S3#
CDR-H2
Residues 50-66
WINTYTGKPTYAQKFQG
36 VH
of SEQ ID
NO.: 1059
J662M2S3#
CDR-H3
Residues 99-112
KLFTTMDVTDNAMD
36 VH
of SEQ ID
NO.: 1059
J662M2S3#
1060
EVQLVQSGAEVKKPGASVKVSCKASGYTFA
45 VH
NYGIIWVRQAPGQGLEWMGWINTYTGKPTY
AQKFQGRVTMTTDTSTSTAYMELSSLRSED
TAVYYCARKLFTTMDVTDNAMDYWGQGTTV
TVSS
J662M2S3#
CDR-H1
Residues 31-35
NYGII
45 VH
of SEQ ID
NO.: 1060
J662M2S3#
CDR-H2
Residues 50-66
WINTYTGKPTYAQKFQG
45 VH
of SEQ ID
NO.: 1060
J662M2S3#
CDR-H3
Residues 99-112
KLFTTMDVTDNAMD
45 VH
of SEQ ID
NO.: 1060
J662M2S3#
1061
EVQLVQSGAEVKKPGASVKVSCKASGYTFS
58 VH
NYGINWVRQAPGQGLEWMGWINTYTGQPSY
AQKFQGRVTMTTDTSTSTAYMELSSLRSED
TAVYYCARKLFKTEAVTDYAMDYWGQGTTV
TVSS
J662M2S3#
CDR-H1
Residues 31-35
NYGIN
58 VH
of SEQ ID
NO.: 1061
J662M2S3#
CDR-H2
Residues 50-66
WINTYTGQPSYAQKFQG
58 VH
of SEQ ID
NO.: 1061
J662M2S3#
CDR-H3
Residues 99-112
KLFKTEAVTDYAMD
58 VH
of SEQ ID
NO.: 1061
J662M2S3#
1062
EVQLVQSGAEVKKPGASVKVSCKASGYTFN
72 VH
NYGIIWVRQAPGQGLEWMGWINTYSGKPTY
AQKFQGRVTMTTDTSTSTAYMELSSLRSED
TAVYFCARKLFTTMDVTDNAMDYWGQGTTV
TVSS
J662M2S3#
CDR-H1
Residues 31-35
NYGII
72 VH
of SEQ ID
NO.: 1062
J662M2S3#
CDR-H2
Residues 50-66
WINTYSGKPTYAQKFQG
72 VH
of SEQ ID
NO.: 1062
J662M2S3#
CDR-H3
Residues 99-112
KLFTTMDVTDNAMD
72 VH
of SEQ ID
NO.: 1062
TABLE 31
Individual hMAK-199 clones VL sequences
Protein
Sequence
region
SEQ ID NO:
123456789012345678901234567890
J662M2S3#
1063
DIQMTQSPSSLSASVGDRVTITCRASQDIS
10 VL
QYLNWYQQKPGKAPKLLIYYTSRLQSGVPS
RFSGSGSGTDFTLTISSLQPEDFATYFCQQ
GNTWPPTFGQGTKLEIK
J662M2S3#10
CDR-L1
Residues 24-34
RASQDISQYLN
VL
of SEQ ID
NO.: 1063
J662M2S3#10
CDR-L2
Residues 50-56
YTSRLQS
VL
of SEQ ID
NO.: 1063
J662M2S3#10
CDR-L3
Residues 89-97
QQGNTWPPT
VL
of SEQ
ID NO.: 1063
J662M2S3#13
1064
DIQMTQSPSSLSASVGDRVTITCRASQDIS
VL
NYLNWYQQKPGKAPKLLIYYTSRLQSGVPS
RFSGSGSGTDYTLTISSLQPEDFATYFCQQ
GNSWPPTFGQGTKLEIK
J662M2S3#13
CDR-L1
Residues 24-34
RASQDISNYLN
VL
of SEQ ID
NO.: 1064
J662M2S3#13
CDR-L2
Residues 50-56
YTSRLQS
VL
of SEQ ID
NO.: 1064
J662M2S3#13
CDR-L3
Residues 89-97
QQGNSWPPT
VL
of SEQ
ID NO.: 1064
J662M2S3#15
1065
DIQMTQSPSSLSASVGDRVTITCRASQDIY
VL
NYLNWYQQKPGKAPKLLIYYTSRLQSGVPS
RFSGSGSGTDYTLTISSLQPEDFATYFCQQ
GNTQPPTFGQGTKLEIK
J662M2S3#15
CDR-L1
Residues 24-34
RASQDIYNYLN
VL
of SEQ ID
NO.: 1065
J662M2S3#15
CDR-L2
Residues 50-56
YTSRLQS
VL
of SEQ ID
NO.: 1065
J662M2S3#15
CDR-L3
Residues 89-97
QQGNTQPPT
VL
of SEQ
ID NO.: 1065
J662M2S3#16
1066
DIQMTQSPSSLSASVGDRVTITCRASQDIE
VL
NYLNWYQQKPGKAPKLLIYYTSRLQSGVPS
RFSGSGSGTDFTLTISSLQPEDFATYFCQQ
GNTQPPTFGQGTKLEIK
J662M2S3#16
CDR-L1
Residues 24-34
RASQDIENYLN
VL
of SEQ ID
NO.: 1066
J662M2S3#16
CDR-L2
Residues 50-56
YTSRLQS
VL
of SEQ ID
NO.: 1066
J662M2S3#16
CDR-L3
Residues 89-97
QQGNTQPPT
VL
of SEQ
ID NO.: 1066
J662M2S3#21
1067
DIQMTQSPSSLSASVGDRVTITCRASQDIS
VL
NYLNWYQQKPGKAPKLLIYYTSRLQSGVPS
RFSGSGSGTDYTLTISSLQPEDFATYFCQQ
GNTWPPTFGQGTKLEIK
J662M2S3#21
CDR-L1
Residues 24-34
RASQDISNYLN
VL
of SEQ ID
NO.: 1067
J662M2S3#21
CDR-L2
Residues 50-56
YTSRLQS
VL
of SEQ ID
NO.: 1067
J662M2S3#21
CDR-L3
Residues 89-97
QQGNTWPPT
VL
of SEQ
ID NO.: 1067
J662M2S3#34
1068
DIQMTQSPSSLSASVGDRVTITCRASQDIY
VL
DVLNWYQQKPGKAPKLLIYYASRLQSGVPS
RFSGSGSGTDFTLTISSLQPEDFATYYCQQ
GITLPPTFGQGTKLEIK
J662M2S3#34
CDR-L1
Residues 24-34
RASQDIYDVLN
VL
of SEQ ID
NO.: 1068
J662M2S3#34
CDR-L2
Residues 50-56
YASRLQS
4 VL
of SEQ ID
NO.: 1068
J662M2S3#34
CDR-L3
Residues 89-97
QQGITLPPT
VL
of SEQ
ID NO.: 1068
J662M2S3#36
1069
DIQMTQSPSSLSASVGDRVTITCRASQDIS
VL
NYLNWYQQKPGKAPKLLIYYTSRLQSGVPS
RFSGSGSGTDYTLTISSLQPEDFATYFCQQ
GNTWPPTFGQGTKLEIK
J662M2S3#36
CDR-L1
Residues 24-34
RASQDISNYLN
VL
of SEQ ID
NO.: 1069
J662M2S3#36
CDR-L2
Residues 50-56
YTSRLQS
VL
of SEQ ID
NO.: 1069
J662M2S3#36
CDR-L3
Residues 89-97
QQGNTWPPT
VL
of SEQ
ID NO.: 1069
J662M2S3#45
1070
DIQMTQSPSSLSASVGDRVTITCRASQDIS
VL
QYLNWYQQKPGKAPKLLIYYTSRLQSGVPS
RFSGSGSGTDFTLTISSLQPEDFATYFCQQ
GNTWPPTFGQGTKLEIK
J662M2S3#45
CDR-L1
Residues 24-34
RASQDISQYLN
VL
of SEQ ID
NO.: 1070
J662M2S3#45
CDR-L2
Residues 50-56
YTSRLQS
VL
of SEQ ID
NO.: 1070
J662M2S3#45
CDR-L3
Residues 89-97
QQGNTWPPT
VL
of SEQ
ID NO.: 1070
J662M2S3#58
1071
DIQMTQSPSSLSASVGDRVTITCRASQNIY
VL
NVLNWYQQKPGKAPKLLIYYASRLQSGVPS
RFSGSGSGTDFTLTISSLQPEDFATYFCQQ
GNTMPPTFGQGTKLEIK
J662M2S3#58
CDR-L1
Residues 24-34
RASQNIYNVLN
VL
of SEQ ID
NO.: 1071
J662M2S3#58
CDR-L2
Residues 50-56
YASRLQS
VL
of SEQ ID
NO.: 1071
J662M2S3#58
CDR-L3
Residues 89-97
QQGNTMPPT
VL
of SEQ
ID NO.: 1071
J662M2S3#72
1072
DIQMTQSPSSLSASVGDRVTITCRASQDIS
VL
NFLNWYQQKPGKAPKLLIYYTSRLQSGVPS
RFSGSGSGTDYTLTISSLQPEDFATYFCQQ
GNTQPPTFGQGTKLEIK
J662M2S3#72
CDR-L1
Residues 24-34
RASQDISNFLN
VL
of SEQ ID
NO.: 1072
J662M2S3#72
CDR-L2
Residues 50-56
YTSRLQS
VL
of SEQ ID
NO.: 1072
J662M2S3#72
CDR-L3
Residues 89-97
QQGNTQPPT
VL
of SEQ
ID NO.: 1072
TABLE 32
hMAK199 affinity matured scFv clones converted to full length IgG
ScFv
Full length IgG
clone name
HC plasmid
LC plasmid
(protein) name
J662M2S3#10
pHybE-hCg1,z,non-a V2
pHybE-hCk V3 J662
hMAK199-AM1
J662M2S3#10
M2S3#10
J662M2S3#13
pHybE-hCg1,z,non-a V2
pHybE-hCk V3 J662
hMAK199-AM2
J662M2S3#13
M2S3#13
J662M2S3#15
pHybE-hCg1,z,non-a V2
pHybE-hCk V3 J662
hMAK199-AM3
J662M2S3#15
M2S3#15
J662M2S3#16
pHybE-hCg1,z,non-a V2
pHybE-hCk V3 J662
hMAK199-AM4
J662M2S3#16
M2S3#16
J662M2S3#21
pHybE-hCg1,z,non-a V2
pHybE-hCk V3 J662
hMAK199-AM5
J662M2S3#21
M2S3#21
J662M2S3#34
pHybE-hCg1,z,non-a V2
pHybE-hCk V3 J662
hMAK199-AM6
J662M2S3#34
M2S3#34
J662M2S3#36
pHybE-hCg1,z,non-a V2
pHybE-hCk V3 J662
hMAK199-AM7
J662M2S3#36
M2S3#36
J662M2S3#45
pHybE-hCg1,z,non-a V2
pHybE-hCk V3 J662
hMAK199-AM8
J662M2S3#45
M2S3#45
J662M2S3#58
pHybE-hCg1,z,non-a V2
pHybE-hCk V3 J662
hMAK199-AM9
J662M2S3#58
M2S3#58
J662M2S3#72
pHybE-hCg1,z,non-a V2
pHybE-hCk V3 J662
hMAK199-AM10
J662M2S3#72
M2S3#72
TABLE 33
hMAK199 affinity matured full length IgG
IgG Name
EC50 in hTNFa ELISA(nM)
hMAK199-AM1
0.016
hMAK199-AM2
0.016
hMAK199-AM3
0.019
hMAK199-AM4
0.050
hMAK199-AM5
0.078
hMAK199-AM6
0.035
hMAK199-AM7
0.100
hMAK199-AM8
0.219
hMAK199-AM9
0.032
hMAK199-AM10
0.014
TABLE 34
hu TNF neutralization
rhesus TNF neutralization
IgG Name
IC50 (nM)
IC50 (nM)
hMAK199-AM1
0.054
0.012
hMAK199-AM2
0.029
0.010
hMAK199-AM3
0.051
0.019
hMAK199-AM4
0.028
0.005
hMAK199-AM5
0.087
0.020
hMAK199-AM6
0.033
0.004
hMAK199-AM7
0.095
0.051
hMAK199-AM8
0.247
0.204
hMAK199-AM9
0.163
0.089
hMAK199-AM10
0.048
0.034
TABLE 35
Reagent for Biacore Analyses
Antigen
Vendor Designation
Vendor
Catalog #
TNFα
Recombinant Human TNF-
R&D
210-TA
α/TNFSF1A
systems
BIACORE Methods:
The BIACORE assay (Biacore, Inc. Piscataway, N.J.) determines the affinity of binding proteins with kinetic measurements of on-rate and off-rate constants. Binding of binding proteins to a target antigen (for example, a purified recombinant target antigen) is determined by surface plasmon resonance-based measurements with a Biacore® 1000 or 3000 instrument (Biacore® AB, Uppsala, Sweden) using running HBS-EP (10 mM HEPES [pH 7.4], 150 mM NaCl, 3 mM EDTA, and 0.005% surfactant P20) at 25° C. All chemicals are obtained from Biacore® AB (Uppsala, Sweden) or otherwise from a different source as described in the text. For example, approximately 5000 RU of goat anti-mouse IgG, (Fcγ), fragment specific polyclonal antibody (Pierce Biotechnology Inc, Rockford, Ill., US) diluted in 10 mM sodium acetate (pH 4.5) is directly immobilized across a CM5 research grade biosensor chip using a standard amine coupling kit according to manufacturer's instructions and procedures at 25 μg/ml. Unreacted moieties on the biosensor surface are blocked with ethanolamine. Modified carboxymethyl dextran surface in flowcell 2 and 4 is used as a reaction surface. Unmodified carboxymethyl dextran without goat anti-mouse IgG in flow cell 1 and 3 is used as the reference surface. For kinetic analysis, rate equations derived from the 1:1 Langmuir binding model are fitted simultaneously to association and dissociation phases of all eight injections (using global fit analysis) with the use of Biaevaluation 4.0.1 software. Purified antibodies are diluted in HEPES-buffered saline for capture across goat anti-mouse IgG specific reaction surfaces. Antibodies to be captured as a ligand (25 μg/ml) are injected over reaction matrices at a flow rate of 5 μl/minute. The association and dissociation rate constants, kon (M−1s−1) and koff (s−1), are determined under a continuous flow rate of 25 μl/minute. Rate constants are derived by making kinetic binding measurements at different antigen concentrations ranging from 10-200 nM. The equilibrium dissociation constant (M) of the reaction between antibodies and the target antigen is then calculated from the kinetic rate constants by the following formula: KD=koff/kon. Binding is recorded as a function of time and kinetic rate constants are calculated. In this assay, on-rates as fast as 106 M−1s−1 and off-rates as slow as 10−6 s−1 can be measured.
The binding proteins herein are expected to have beneficial properties in this regard, including high affinity, slow off rate, and high neutralizing capacity.
L929 cells are grown to a semi-confluent density and harvested using 0.25% trypsin (Gibco #25300). The cells are washed with PBS, counted and resuspended at 1E6 cells/mL in assay media containing 4 μg/mL actinomycin D. The cells are seeded in a 96-well plate (Costar #3599) at a volume of 100 μL and 5E4 cells/well. The binding proteins and control IgG are diluted to a 4× concentration in assay media and serial 1:4 dilutions are performed. The huTNF-α is diluted to 400 pg/mL in assay media. Binding protein sample (200 μL) is added to the huTNF-α (200 μL) in a 1:2 dilution scheme and allowed to incubate for 0.5 hour at room temperature.
The binding protein/human TNF-α solution is added to the plated cells at 100 μL for a final concentration of 100 pg/mL huTNF-α and 150 nM-0.0001 nM binding protein. The plates are incubated for 20 hours at 37° C., 5% CO2. To quantitate viability, 100 μL is removed from the wells and 10 μL of WST-1 reagent (Roche cat #11644807001) is added. Plates are incubated under assay conditions for 3.5 hours. The plates are read at OD 420-600 nm on a Spectromax 190 ELISA plate reader.
The binding proteins herein are expected to have beneficial properties in this regard, including high affinity, slow off rate, and high neutralizing capacity.
A patient requiring treatment with a TNF-α binding protein may have a disease with immune and inflammatory elements, such as autoimmune diseases, particularly those assocated with inflammation, including Crohn's disease, psoriasis (including plaque psoriasis), arthritis (including rheumatoid arthritis, psoratic arthritis, osteoarthritis, or juvenile idiopathic arthritis), multiple sclerosis, and ankylosing spondylitis. Therefore, the binding proteins herein may be used to treat these disorders.
Administration of the TNF-α binding protein may occur by subcutaneous injection. If the patient has rheumatoid arthritis, psoratic arthritis, or ankylosing spondyitis, the patient may receive 40 mg every other week as a starting dose and 40 mg every week, if necessary to achieve treatment goals. If the patient has juvenile idiopathic arthritis and weighs from 15 kg to <30 kg, the patient may receive 20 mg every other week, and if ≧30 kg, 40 mg every other week. If the patient has Crohn's disease, the patient may receive an initial dose of 160 mg (four 40 mg injections in one day or two 40 mg injections per day for two consecutive days) followed by 80 mg two weeks later, and another two weeks later begin a maintenance dose of 40 mg every other week. If the patient has plaque psoriasis, the patient may receive an 80 mg initial dose, followed by 40 mg every other week starting one week after initial dose.
The binding protein may be provided in a single-use prefilled pen (40 mg/0.8 mL), a single-use prefilled glass syringe (40 mg/0.8 mL or 20 mg/0.4 mL).
The contents of all cited references (including literature references, patents, patent applications, and websites) that are cited throughout this application are hereby expressly incorporated by reference in their entirety, as are the references cited therein. The practice disclosed herein will employ, unless otherwise indicated, conventional techniques of immunology, molecular biology and cell biology, which are well known in the art.
The invention may be embodied in other specific forms without departing from the spirit or essential characteristics thereof. The foregoing embodiments are therefore to be considered in all respects illustrative rather than limiting of the invention described herein. Scope of the invention is thus indicated by the appended claims rather than by the foregoing description, and all changes that come within the meaning and range of equivalency of the claims are therefore intended to be embraced herein.
Hsieh, Chung-ming, Kutskova, Yuliya, Zhong, Suju, Clabbers, Anca, Benatuil, Lorenzo, Perez, Jennifer, Memmott, John, Goodreau, Carrie
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| Dec 03 2012 | KUTSKOVA, YULIYA | AbbVie Inc | ASSIGNMENT OF ASSIGNORS INTEREST SEE DOCUMENT FOR DETAILS | 029946 | /0042 | |
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